Recombinant ribonuclease proteins

ABSTRACT

PCT No. PCT/US97/02588 Sec. 371 Date Feb. 19, 1998 Sec. 102(e) Date Feb. 19, 1998 PCT Filed Feb. 19, 1997 PCT Pub. No. WO97/31116 PCT Pub. Date Aug. 28, 1997The invention relates to ribonucleases derived from a native ribonuclease found in the oocytes of Rana pipiens. Various humanized and recombinant forms of these molecules are described as well as uses for them.

This is a National Stage Application of PCT/US97/02588, filed Feb. 19,1997 and a continuation of Provisional Application No. 06/011,800 filedFeb. 21, 1996.

FIELD OF THE INVENTION

This invention relates to the production of ribonuclease molecules whichare toxic to cells of interest.

BACKGROUND OF THE INVENTION

Ribonucleases such as ribonuclease A ("RNase A") and their cytotoxicitytoward tumor cells are well documented from studies performed in the1960s and 1970s and reviewed in Roth, J., 1963, Cancer Res. 23:657-666.Human serum was also discovered to contain several RNases (Reddi, E.,1975, Biochem. Biophys. Res. Commun. 67:110-118, Blank et al., Humanbody fluid ribonucleases: detection, interrelationships and significance1-203-209 (IRL Press, London, 1981)) that are expressed in a tissuespecific manner. The proteins involved in the host defense activity ofthe eosinophil are homologous to RNases and express RNase activity(Gleich et al., 1986, Proc. Natl. Acad. Sci., USA 83:3146-3150; Slifmanet al., 1986, J. Immunol, 137:2913-2917). Thus, human serum RNases werebelieved to also have host defense activities.

Further to these early studies was the discovery that an anti-tumorprotein from oocytes of Rana pipiens has homology to RNase A (Ardelt etal., 1991, J. Biol. Chem. 266:245-251). This protein has been termedONCONASE®, Alfacell Corporation, N.J. See also e.g., Darzynkiewicz etal. (1988) Cell Tissue Kinet. 21, 169-182, Mikulski et al. (1990) CellTissue Kinet. 23, 237-246. This protein is also described in U.S. Pat.No. 4,888,172. Phase I and Phase I/II clinical trials of ONCONASE® as asingle therapeutic agent in patients with a variety of solid tumors(Mikulski et al. (1993) Int. J. of Oncology 3, 57-64) or combined withtamoxifen in patients with advanced pancreatic carcinoma have recentlybeen completed (Chun et al. (1995) Proc Amer Soc Clin Oncol 14 No. 157,210). Conjugation of ONCONASE® to cell-type-specific ligands increasedits potency towards tumor cells (Rybak et al. (1993) Drug Delivery 1,3-10). Taken together, these results indicate that ONCONASE® hasproperties that are advantageous for the generation of a potentselective cell killing agent.

However, since this is not a human-derived protein, it is prone tostimulating undesirable immune responses when used in humans. Thus, itwould be desirable to retain the potent cytotoxic properties of thismolecule while reducing its immunogenicity in humans. Further, it wouldbe desirable to produce derivations of this molecule recombinantly sothat it may be better chemically conjugated or recombinantly joined toother molecules for targeting to specific cells. Until the inventiondescribed herein, it has proven difficult to recombinantly express anactive cytotoxic molecule related to ONCONASE®. Though it was thoughtthat the methionine-glutamic acid amino terminal end of the recombinantmolecule prohibited the molecule from having significant enzymaticactivity, a means to solve this problem has not been forthcoming untilthe invention herein.

Further, although advances in protein design techniques promise toalleviate some of the immunogenicity associated with the antibodyportion of immunotoxins (Bird et al., 1988, Science 242:423; Huston etal., 1988, Proc Natl Acad Sci USA 85:5879; Ward et al., 1989, Nature341:544), no solution has been forthcoming for the immunogenicity of thetoxin portion other than immunosuppression of the patients (Khazaeli etal., 1988, Proceedings of AACR 29:418). Thus, there is a continuing needfor methods and compositions that would reduce the immunogenicity of theRana pipiens-derived toxic moiety.

Non-cytotoxic human members of the RNase A superfamily linked to tumorassociated antigens by chemical (Rybak et al.(1991) J. Biol. Chem 266,1202-21207, Newton et al. (1992) J. Biol. Chem. 267, 19572-19578) orrecombinant means (Rybak et al. Proc. Natl. Acad. Sci. U.S.A. 89, 3165,Newton et al. (1994) J Biol Chem. 269, 26739-26745 have been shown tooffer a strategy for selectively killing tumor cells with lessimmunogenicity than current strategies employing plant and bacterialtoxins Rybak, S. M. & Youle, R. J. (1991) Immunol. and Allergy Clinicsof North America 11:2, 359-380. Human-derived ribonucleases of interestinclude eosinophil-derived neurotoxin (EDN) and angiogenin.

SUMMARY OF THE INVENTION

We have discovered how to construct RNases which are highly cytotoxicand which are modifications of the native ONCONASE® (nOnc). When thenOnc was expressed recombinantly it was not found to have significantcytotoxicity. Our modified versions (rOnc), however, are highlycytotoxic and otherwise retain the advantages of the native ONCONASE®molecules, while in some cases they also have increased cytotoxicproperties. The rOnc molecules may be used alone or conveniently used toform chemical conjugates, as well as to form targeted recombinantimmunofusions. These rOnc molecules can be used to decrease tumor cellgrowth. An effective recombinant form of nOnc advantageously permits therecombinant molecule to be fused to other therapeutic or targetingmolecules of interest recombinantly. Further, the rOnc molecule can bemodified to enhance cytotoxicity as will be seen below. Our nOnc-derivedmolecules are also desirable because nOnc is a unique ribonuclease inthat it can be administered alone directly to patients to decrease andinhibit tumor cell growth without the use of a targeting agent.

The present invention also includes methods of selectively killing cellsusing a rOnc joined to a ligand to create a selective cytotoxic reagentof the present invention. The method comprises contacting the cells tobe killed with a cytotoxic reagent of the present invention having aligand binding moiety that specifically delivers the reagent to thecells to be killed. This method of the present invention may be used forcell separation in vitro by selectively killing unwanted types of cells,for example, in bone marrow prior to transplantation into a patientundergoing marrow ablation by radiation, or for killing leukemia cellsor T-cells that would cause graft-versus-host disease. The toxins canalso be used to selectively kill unwanted cells in culture.

Humanized versions of our rOnc molecules are also described which graftportions of mammalian or human-derived RNases such as angiogenin orhuman eosinophil derived neurotoxin (EDN) to the rOnc-derived molecules.A preferred embodiment of the invention is a molecule where the aminoterminal end of EDN is placed onto the amino terminal end of the rOncmolecules. The surprising properties of these hybrid proteins withregard to ribonuclease activity and in vitro anti-tumor effects aredescribed.

BRIEF DESCRIPTION OF THE DRAWINGS FIGURE LEGENDS

FIG. 1 shows the deduced amino acid sequence of the Rana clone 9 (SEQ IDNO:2), described below and sequence alignment with the amino acidsequence of nOnc (SEQ ID NO:1). The bold print indicates identicalresidues between nOnc and Rana clone 9. The dots indicate missing aminoacids in the PCR clone.

FIGS. 2A and 2B show the configuration of the DNA constructs exemplifiedin the examples. The PCR product obtained from Rana pipiens DNA isidentified as Rana 9. The N-and C-termini are synthetically filled inand identified as Onc in the constructs encoding [Met-(-1)]rOnc or EDNfor the N-terminal EDN/Onc hybrid. Corresponding amino acid residues areindicated below each construct. FIG. 2B shows the sequence alignment ofthe N-terminal sequences of nOnc (SEQ ID NO:3), rEDN (SEQ ID NO:4),[Met-(-1)]rOnc containing a Gly (G) instead of Asp in position 20 (SEQID NO:5), rEDN.sub.(1-21) rOnc with an Asp in amino acid position 26(SEQ ID NO:6) and rEDN.sub.(1-21) rOncG26 with a Gly in position 26 (SEQID NO:7). Bold letters indicate conserved residues, capital letters showthe sequence deduced from Rana clone 9.

FIGS. 3A-3D show the inhibition of protein synthesis in human tumorcells by nOnc, rEDN, [Met-(-1)]rOnc or hybrid proteins. Cells (10⁴) wereplated in individual 96-well microtiter culture plates and treated withvarying concentrations of each agent for 48 h. Cell viability wasdetermined as described in the Example Section below. Results from morethan one individual experiment were combined to yield the mean datapoints. Standard errors of the means, when they are greater than thesymbol, are shown. Cell lines: ACHN, renal cancer (FIG. 3A); MDA-MB-231(FIG. 3B) and HS 578T (FIG. 3D), breast cancer; SF-539 (FIG. 3C), CNS,cancer. EDN (open triangles); nOnc (open squares); [Met-(-1)]rOnc,(solid triangles); rEDN.sub.(1-21) rOnc, (open circles); rEDN.sub.(1-21)rOncG26 (solid circles).

FIG. 4 shows a sequence alignment of some members of the RNase Asuperfamily: Frog lectin is from Rana catesbeiana, ONCONASE®, EDN, ECP(human eosinophil cationic protein), Ang is bovine angiogenin, Seminalis bovine seminal RNase, and RNase A is bovine pancreatic RNase A (SEQID NOs:8, 1 and 9-13, respectively). Amino acids conserved in allmembers are capitalized, and active site residues H12, K41, and H119(RNase A numbering) are marked with an asterisk.

FIG. 5 shows the inhibition of protein synthesis by MetSerOnc andMetSer- or MetGlu-OncFvs. The cytotoxic effect of the single chainantibody rOnc fusion proteins; E6FB[Met-(-1)]SerrOnc (closed circles),[Met-(-1)]SerrOncAngFBE6 (open squares) and [Met-(-1)]GlurOncFBE6(closed squares) were compared to the non-targeted recombinant protein,[Met-(-1)]SerrOnc (open circles), by determining inhibition of proteinsynthesis in SF 539 cells. Cells were plated into 96-well microtiterplates in Dulbecco's minimum essential medium supplemented with 10%heat-inactivated fetal bovine serum. Additions were made in a totalvolume of 10 μl and the plates were incubated at 37° for 3 days.Phosphate buffered saline containing 0.1 mCi of [¹⁴ C]leucine was addedfor 2-4 h and the cells were harvested onto glass fiber filters using aPHD cell harvester, washed with water, dried with ethanol and counted.The results were expressed as per cent of [¹⁴ C]leucine incorporation inthe mock-treated wells.

FIGS. 6A and 6B show inhibition of protein synthesis in an assay asdescribed for FIGS. 3A-3D using cell line SF539, human glioma cells andrOnc fusion proteins designated MetLysTryrOnc (open circles, FIG. 6A);MetAlaAlaTyrOnc (closed circles, FIG. 6A); and rOnc fusion proteins withsignal peptides, MetKDELSerrOnc (open circles, FIG. 6B) andMetNLSSerrOnc (closed circles FIG. 6B).

FIG. 7 shows inhibition of protein synthesis in an assay as describedfor FIGS. 3A-3D using cell line SF539, human glioma cells and comparingthree fusion proteins corresponding to MetSerOnc (SEQ ID NO:39 with aMet-Ser amino terminal end): MetSerOnc (closed circles), MetSerOncC4(MetSerOnc with a Cys at amino acid position 5 of SEQ ID NO:39, closedsquares) and MetSerOncC72 (MetSerOnc with a Cys at amino acid position73 of SEQ ID NO:39, open circles).

DETAILED DESCRIPTION

This invention provides highly active and cytotoxic ribonucleasemolecules which can be used to selectively kill and target cells,particularly tumor cells. In some embodiments the molecules are designedto incorporate sequences from human derived ribonucleases which are alsohighly active and cytotoxic, but which have the further advantage inthat they are less immunogenic in humans. The rOnc molecules of thepresent invention are those which are recombinant nOnc-derivedsequences.

The nOnc molecule has an amino acid sequence set forth in SEQ ID NO:1.Bovine pancreatic RNase A has an amino acid sequence set forth in SEQ IDNO:13. Unless otherwise indicated, the amino acid sequence positionsdescribed herein use as a frame of reference the bovine pancreatic RNaseA sequence in SEQ ID NO:13 as this is the reference sequence commonlyused in the RNase field. It should be understood that such positiondesignations do not indicate the number of amino acids in the claimedmolecule per se, but indicate where in the claimed molecule the residueoccurs when the claimed molecule sequence is aligned with bovine RNase.

The rOnc molecules described and claimed herein will preferably havecysteine residues at amino acid positions corresponding to amino acidpositions 26, 40, 58, 84, 95 and 110; a lysine at position 41 and ahistidine at position 119 with reference to the bovine RNase A, SEQ IDNO:13 (such positions correspond to amino acid position numbers 19, 30,48, 68, 75 and 90 and 87 and 104 of the nOnc sequence respectively setout in SEQ ID NO:1).

The rOnc molecules of this invention are those that have measurableribonuclease activity, as defined below. The ribonucleases will alsohave (a) an amino terminal end beginning with a methionine which isfollowed by any amino acid other than glutamic acid (Glu); (b) acysteine at amino acid positions 26, 40, 58, 84, 95 and 110; a lysine atposition 41 and a histidine at position 119, such positions beingdetermined with reference to those in the amino acid sequence of bovineRNase A (SEQ ID NO:13), and (c) an nOnc-derived amino acid sequence.

Preferably, the rOnc molecules will have an amino terminal end selectedfrom the group consisting of:

Met-Ala;

Met-Ala-Ala-Ser;

Met-Arg;

Met-(J);

Met-Lys-(J);

Met-Arg-(J);

Met-Lys;

Met-Lys-Pro;

Met-Lys-(J)-Pro (SEQ ID NO:14);

Met-Lys-Pro-(J) (SEQ ID NO:15);

Met-Asn;

Met-Gln;

Met-Asn-(J);

Met-Gln-(J);

Met-Asn-(J)-Pro (SEQ ID NO:16);

Met-(J)-Lys;

Met-(J)-Lys-Pro (SEQ ID NO:17); and

Met-(J)-Pro-Lys (SEQ ID NO:18);

where (J) is Ser, Tyr or Thr.

Further, it is preferred that the rOnc molecules be modified so that theaspartic acid of amino acid position 2 of nOnc (position 4 withreference to the sequence of bovine RNase A) is deleted or replaced byAla or Asn.

In alternative forms of the rOnc molecules, the molecules will employ anamino terminal end encoded by a sequence derived from the amino terminalend of EDN followed by a sequence from rOnc. In such forms, it ispreferred that the amino acid sequence is one selected from the groupconsisting of those sequences substantially identical to those of aformula:

    Met(-1)EDN.sub.(1-m) Onc.sub.(n-104)

wherein Met(-1) refers to an amino terminal residue of Met; whereinEDN.sub.(1-m) refers to a contiguous sequence of amino acids of a lengthbeginning at amino acid position 1 of EDN (SEQ ID NO:9) and continuingto and including amino acid position "m" of EDN; wherein Onc.sub.(n-104)refers to a sequence of contiguous amino acids beginning at amino acidposition "n" and continuing to and including amino acid position 104 asset out in SEQ ID NO:1; such that:

when m is 21, n is 16 or 17;

when m is 22, n is 17;

when m is 20, n is 16;

when m is 19, n is 15;

when m is 18, n is 14;

when m is 17, n is 12 or 13;

when m is 16, n is 11, 12, 13 or 14;

when m is 15, n is 10;

when m is 14, n is 9;

when m is 13, n is 8; and

when m is 5, n is 1.

In other alternative embodiments, the rOnc molecule will be fused at thecarboxyl end to a sequence from angiogenin, such as the sequenceexemplified in SEQ ID NO:11 or that at amino acid positions 101 to 107of SEQ ID NO:20. The nucleic acid sequence for human angiogenin is knownand is set out in U.S. patent application Ser. No. 08/125,462.

Preferred rOnc nucleic acid sequences are those that encode preferredrOnc amino acid sequences which are substantially identical to those inSEQ ID NOs:20, 22, 24, 26, 28 and 30 (corresponding nucleic acidsequences are set out in SEQ ID NOs:19, 21, 23, 25, 27 and 29,respectively). Most preferred rOnc amino acid sequences are those thatare substantially identical to those set forth in SEQ ID NOs:20, 22, 24and 26. Their corresponding nucleic acid sequences are also preferredand are set out in SEQ ID NOs:19, 21, 23 and 25, includingconservatively modified variants thereof. The most preferred sequenceincludes SEQ ID NO:22, one which employs an amino terminal endcomprising 1 to 21 (typically 21) amino acids of the amino terminal endof EDN grafted on to 16 to 104 amino acids of the nOnc sequence, withamino acid residue 20 in nOnc (Asp) being replaced with Gly. PreferredrOnc sequences further will contain optionally a Cys at a positioncorresponding to amino acid position 5, or 73 or Ala at amino acidposition 88 in place of Cys with reference to SEQ ID NO:39.

Comparisons of the rOnc sequences provided here can be made to describedsequences in the pancreatic RNase A superfamily. Many of such membersare known and include, but are not limited to, frog lectin from Ranacatesbeiana (Titani et al., Biochemistry 26:2189 (1987)); ONCONASE®(Ardelt, W. et al., J. Biol. Chem. 266:245 (1991)); eosinophil derivedneurotoxin (EDN) (Rosenberg et al., supra); human eosinophil cationicprotein (ECP) (Rosenberg et al., J. Exp. Med. 170:163 (1989));angiogenin (Ang) (Fett, J. W. et al., Biochemistry 24:5480 (1985));bovine seminal RNase (Preuss et al., Nuc. Acids. Res. 18:1057 (1990));and bovine pancreatic RNase (Beintama et al., Prog. Biophys. Mol. Biol.51:165 (1988)), references for all such proteins are incorporated byreference herein. Amino acid sequence alignment for such RNases are alsoset out in FIG. 4 and in Youle et al., Crit. Rev. Ther. Drug. CarrierSystems 10:1-28 (1993) and in U.S. patent application Ser. No.08/125,462, which is incorporated by reference herein.

DEFINITIONS

Unless defined otherwise herein, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention belongs. Singleton et al.(1994) Dictionary of Microbiology and Molecular Biology, second edition,John Wiley and Sons (New York), and Hale and Marham (1991) The HarperCollins Dictionary of Biology, Harper Perennial, NY provide one of skillwith a general dictionary of many of the terms used in this invention.Although any methods and materials similar or equivalent to thosedescribed herein can be used in the practice or testing of the presentinvention, the preferred methods and materials are described. Forpurposes of the present invention, the following terms are definedbelow.

Amino acids may be referred to herein by either their commonly knownthree letter symbols or by the one-letter symbols recommended by theIUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise,may be referred to by their commonly accepted single-letter codes.

The terms "measurable ribonuclease activity" or "significantribonuclease activity" refer to a molecule which has an IC₅₀ (ng/ml) ofless than 40 when added to a rabbit reticulocyte lysate assay whereinprotein synthesis is inhibited as measured by the incorporation of [³⁵S]methionine into acid precipitable protein. IC₅₀ is the concentrationof protein necessary to inhibit protein synthesis by 50% in the assay.The lysate assay may be done as described in the Promega lysate assaykit which is commercially available from Promega Corporation, Madison,Wis. Ribonuclease activity using high molecular weight RNA and tRNA isdetermined at 37° C. through the formation of perchloric acid solublenucleotides following published protocols (Newton, D. L., et al. (1996)Biochemistry 35:545-553). With poly(A,C) UpG and poly U, ribonucleaseactivity is assayed according to DePrisco et al., and Libonati andFloridi (DePrisco, R., et al. (1984) Biochimica et Biophysica Acta788:356-363; Libonati, M. et al. (1969) European J. Biochem. 8:81-87).Activity is assayed by measuring the increase with time in absorbance at260 nm. Incubation mixtures (1 ml of 10 mM imidazole, 0.1 M NaCl, pH 6.5or pH 7) contain substrate and appropriate amounts of enzyme solution at25° C. The in vitro translation assay (St. Clair, D. K., et al. (1987)Proc. Natl. Acad. Sci. 84:8330-8334) and the cell viability assays usingthe (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide;Thiazolyl blue) (MMT) (Mossman, T. (1983) J. Immunol. Methods 65:55-63)are performed as previously described (Pearson, J. W., et al. (1991) J.Natl. Cancer Inst. 83:1386-1391).

An "nOnc-derived" amino acid sequence is one that contains at least onestring of six contiguous amino acids which is identical to a contiguoussequence of six amino acids selected from the group of sequencesbeginning at amino acid positions 1 (with Glu replacing pyroGlu), 2, 3,4, 5, 6, 7, 8, 11, 12, 13, 14, 15, 16, 18, 19, 20, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 41, 42, 43, 44, 45, 46, 47,50, 52, 54, 56, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 76, 80, 81, 82, 84, 85, 86, 87, 91, 92, 93, 95, or 96 of thenOnc amino acid sequence (SEQ ID NO:1).

"Conservatively modified variations" of a particular nucleic acidsequence refer to those nucleic acids which encode identical oressentially identical amino acid sequences, or where the nucleic aciddoes not encode an amino acid sequence, to essentially identicalsequences. Because of the degeneracy of the genetic code, a large numberof functionally identical nucleic acids encode any given polypeptide.For instance, the codons GCA, GCC, GCG and GCU all encode the amino acidalanine. Thus, at every position where an alanine is specified by acodon, the codon can be altered to any of the corresponding codonsdescribed without altering the encoded polypeptide. Such nucleic acidvariations are "silent variations," which are one species ofconservatively modified variations. Every nucleic acid sequence hereinwhich encodes a polypeptide also describes every possible silentvariation of the nucleic acid. One of skill will recognize that eachcodon in a nucleic acid (except AUG, which is ordinarily the only codonfor methionine) can be modified to yield a functionally identicalmolecule. Accordingly, each silent variation of a nucleic acid whichencodes a polypeptide is implicit in each described sequence.Furthermore, one of skill will recognize that individual substitutions,deletions or additions which alter, add or delete a single amino acid ora small percentage of amino acids in an encoded sequence are"conservatively modified variations" where the alterations result in thesubstitution of an amino acid with a chemically similar amino acid.Conservative substitution tables providing functionally similar aminoacids are well known in the art. The following six groups each containamino acids that are conservative substitutions for one another:

1) Alanine (A), Serine (S), Threonine (T);

2) Aspartic acid (D), Glutamic acid (E);

3) Asparagine (N), Glutamine (Q);

4) Arginine (R), Lysine (K);

5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and

6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

See also, Creighton (1984) Proteins W. H. Freeman and Company.

The terms "isolated" or "biologically pure" refer to material which issubstantially or essentially free from components which normallyaccompany it as found in its naturally occurring environment. Theisolated material optionally comprises material not found with thematerial in its natural environment. The rOncs described herein areisolated and biologically pure since they are recombinantly produced inthe absence of unrelated Rana pipiens proteins. They may, however,include heterologous cell components, a ligand binding moiety, a labeland the like.

The term "nucleic acid" refers to a deoxyribonucleotide orribonucleotide polymer in either single- or double-stranded form, andunless otherwise limited, encompasses known analogues of naturalnucleotides that hybridize to nucleic acids in a manner similar tonaturally occurring nucleotides. Unless otherwise indicated, aparticular nucleic acid sequence includes the complementary sequencethereof. A nucleic acid encodes another nucleic acid where it is thesame as the specified nucleic acid, or complementary to the specifiednucleic acid.

An "expression vector" includes a recombinant expression cassette whichincludes a nucleic acid which encodes a rOnc polypeptide which can betranscribed and translated by a cell. A recombinant expression cassetteis a nucleic acid construct, generated recombinantly or synthetically,with a series of specified nucleic acid elements which permittranscription of a particular nucleic acid in a target cell. Theexpression vector can be part of a plasmid, virus, or nucleic acidfragment. Typically, the recombinant expression cassette portion of theexpression vector includes a nucleic acid to be transcribed, and apromoter.

The term "recombinant" when used with reference to a protein indicatesthat a cell expresses a peptide or protein encoded by a nucleic acidwhose origin is exogenous to the cell. Recombinant cells can expressgenes that are not found within the native (non-recombinant) form of thecell. Recombinant cells can also express genes found in the native formof the cell wherein the genes are re-introduced into the cell byartificial means, for example under the control of a heterologouspromoter.

The term "subsequence" in the context of a particular nucleic acid orpolypeptide sequence refers to a region of the nucleic acid orpolypeptide equal to or smaller than the particular nucleic acid orpolypeptide. "Stringent hybridization wash conditions" in the context ofnucleic acid hybridization experiments such as Southern and northernhybridizations are sequence dependent, and are different under differentenvironmental parameters. An extensive guide to the hybridization ofnucleic acids is found in Tijssen (1993) Laboratory Techniques inBiochemistry and Molecular Biology--Hybridization with Nucleic AcidProbes Part I, Chapter 2 "Overview of principles of hybridization andthe strategy of nucleic acid probe assays", Elsevier, New York.Generally, highly stringent wash conditions are selected to be about 5°C. lower than the thermal melting point (T_(m)) for the specificsequence at a defined ionic strength and ph. The T_(m) is thetemperature (under defined ionic strength and pH) at which 50% of thetarget sequence hybridizes to a perfectly matched probe. Very stringentconditions are selected to be equal to the T_(m) point for a particularprobe. Nucleic acids which do not hybridize to each other understringent conditions are still substantially identical if thepolypeptides which they encode are substantially identical. This occurs,e.g., when a copy of a nucleic acid is created using the maximum codondegeneracy permitted by the genetic code.

The term "identical" in the context of two nucleic acid or polypeptidesequences refers to the residues in the two sequences which are the samewhen aligned for maximum correspondence over a specified comparisonwindow. When percentage of sequence identity is used in reference toproteins or peptides it is recognized that residue positions which arenot identical often differ by conservative amino acid substitutions,where amino acid residues are substituted for other amino acid residueswith similar chemical properties (e.g. charge or hydrophobicity) andtherefore do not change the functional properties of the molecule. Wheresequences differ in conservative substitutions, the percent sequenceidentity may be adjusted upwards to correct for the conservative natureof the substitution. Means for making this adjustment are well-known tothose of skill in the art. Typically this involves scoring aconservative substitution as a partial rather than a full mismatch,thereby increasing the percentage sequence identity. Thus, for example,where an identical amino acid is given a score of 1 and anon-conservative substitution is given a score of zero, a conservativesubstitution is given a score between zero and 1. The scoring ofconservative substitutions is calculated, e.g., according to thealgorithm of Meyers and Miller, Computer Applic. Biol. Sci., 4: 11-17(1988) e.g., as implemented in the program PC/GENE (Intelligenetics,Mountain View, Calif., U.S.A.).

Methods of alignment of sequences for comparison are well-known in theart. Optimal alignment of sequences for comparison may be conducted bythe local homology algorithm of Smith and Waterman (1981) Adv. Appl.Math. 2: 482; by the homology alignment algorithm of Needleman andWunsch (1970) J. Mol. Biol. 48: 443; by the search for similarity methodof Pearson and Lipman (1988) Proc. Natl. Acad. Sci. USA 85: 2444; bycomputerized implementations of these algorithms (including, but notlimited to CLUSTAL in the PC/Gene program by Intelligenetics, MountainView, Calif., GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin GeneticsSoftware Package, Genetics Computer Group (GCG), 575 Science Dr.,Madison, Wis., U.S.A.); the CLUSTAL program is well described by Higginsand Sharp (1988) Gene, 73: 237-244 and Higgins and Sharp (1989) ComputerApplications in the Biosciences 5: 151-153; Corpet, et al. (1988)Nucleic Acids Research 16, 10881-90; Huang, et al. (1992) ComputerApplications in the Biosciences 8, 155-65, and Pearson, et al. (1994)Methods in Molecular Biology 24, 307-31. Alignment is also oftenperformed by inspection and manual alignment.

The term "substantial identity" or "substantial similarity" in thecontext of a polypeptide indicates that a polypeptide comprises asequence with at least 70% sequence identity to a reference sequence, orpreferably 80%, or more preferably 85% sequence identity to thereference sequence, or most preferably 90% identity over a comparisonwindow of about 10-20 amino acid residues. An indication that twopolypeptide sequences are substantially identical is that one peptide isimmunologically reactive with antibodies raised against the secondpeptide. Thus, a polypeptide is substantially identical to a secondpolypeptide, for example, where the two peptides differ only by aconservative substitution.

One indication that two nucleic acid sequences are substantiallyidentical is that the polypeptide which the first nucleic acid encodesis immunologically cross reactive with the polypeptide encoded by thesecond nucleic acid.

Another indication that two nucleic acid sequences are substantiallyidentical is that the two molecules hybridize to each other understringent conditions. Stringent conditions are sequence dependent andare different under different environmental parameters. Generally,stringent conditions are selected to be about 5° C. to 20° C. lower thanthe thermal melting point (T_(m)) for the specific sequence at a definedionic strength and pH. The T_(m) is the temperature (under defined ionicstrength and pH) at which 50% of the target sequence hybridizes to aperfectly matched probe. However, nucleic acids which do not hybridizeto each other under stringent conditions are still substantiallyidentical if the polypeptides which they encode are substantiallyidentical. This occurs, e.g., when a copy of a nucleic acid is createdusing the maximum codon degeneracy permitted by the genetic code.

The term "specifically deliver" as used herein refers to thepreferential association of a molecule with a cell or tissue bearing aparticular target molecule or marker and not to cells or tissues lackingthat target molecule. It is, of course, recognized that a certain degreeof non-specific interaction may occur between a molecule and anon-target cell or tissue. Nevertheless, specific delivery, may bedistinguished as mediated through specific recognition of the targetmolecule. Typically specific delivery results in a much strongerassociation between the delivered molecule and cells bearing the targetmolecule than between the delivered molecule and cells lacking thetarget molecule. Specific delivery typically results in greater than 2fold, preferably greater than 5 fold, more preferably greater than 10fold and most preferably greater than 100 fold increase in amount ofdelivered molecule (per unit time) to a cell or tissue bearing thetarget molecule as compared to a cell or tissue lacking the targetmolecule or marker.

The term "residue" as used herein refers to an amino acid that isincorporated into a polypeptide. The amino acid may be a naturallyoccurring amino acid and, unless otherwise limited, may encompass knownanalogs of natural amino acids that can function in a similar manner asnaturally occurring amino acids.

A "fusion protein" or when a molecule is "joined" to another refers to achimeric molecule formed by the joining of two or more polypeptidesthrough a peptide bond formed between the amino terminus of onepolypeptide and the carboxyl terminus of another polypeptide. The fusionprotein or the joined molecules may be formed by the chemical couplingof the constituent molecules or it may be expressed as a singlepolypeptide from a nucleic acid sequence encoding a single contiguousfusion protein. A single chain fusion protein is a fusion protein havinga single contiguous polypeptide backbone.

A "ligand" or a "ligand binding moiety", as used herein, refersgenerally to all molecules capable of specifically delivering amolecule, reacting with or otherwise recognizing or binding to areceptor on a target cell. Specifically, examples of ligands include,but are not limited to, antibodies, lymphokines, cytokines, receptorproteins such as CD4 and CD8, solubilized receptor proteins such assoluble CD4, hormones, growth factors, and the like which specificallybind desired target cells.

Making rOnc-derived Nucleic Acids and Polypeptides

Several specific nucleic acids encoding rOnc-derived polypeptides aredescribed herein. These nucleic acids can be made using standardrecombinant or synthetic techniques. Given the nucleic acids of thepresent invention, one of skill can construct a variety of clonescontaining functionally equivalent nucleic acids, such as nucleic acidswhich encode the same polypeptide. Cloning methodologies to accomplishthese ends, and sequencing methods to verify the sequence of nucleicacids are well known in the art. Examples of appropriate cloning andsequencing techniques, and instructions sufficient to direct persons ofskill through many cloning exercises are found in Berger and Kimmel,Guide to Molecular Cloning Techniques, Methods in Enzymology volume 152Academic Press, Inc., San Diego, Calif. (Berger); Sambrook et al. (1989)Molecular Cloning--A Laboratory Manual (2nd ed.) Vol. 1-3; and CurrentProtocols in Molecular Biology, F. M. Ausubel et al., eds., CurrentProtocols, a joint venture between Greene Publishing Associates, Inc.and John Wiley & Sons, Inc., (1994 Supplement) (Ausubel). Productinformation from manufacturers of biological reagents and experimentalequipment also provide information useful in known biological methods.Such manufacturers include the SIGMA chemical company (Saint Louis,Mo.), R&D systems (Minneapolis, Minn.), Pharmacia LKB Biotechnology(Piscataway, N.J.), CLONTECH Laboratories, Inc. (Palo Alto, Calif.),Chem Genes Corp., Aldrich Chemical Company (Milwaukee, Wis.), GlenResearch, Inc., GIBCO BRL Life Technologies, Inc. (Gaithersberg, Md.),Fluka Chemica-Biochemika Analytika (Fluka Chemie AG, Buchs,Switzerland), Invitrogen, San Diego, Calif., and Applied Biosystems(Foster City, Calif.), as well as many other commercial sources known toone of skill.

The nucleic acid compositions of this invention, whether RNA, cDNA,genomic DNA, or a hybrid of the various combinations, are isolated frombiological sources or synthesized in vitro. The nucleic acids of theinvention are present in transformed or transfected cells, intransformed or transfected cell lysates, or in a partially purified orsubstantially pure form.

In vitro amplification techniques suitable for amplifying sequences foruse as molecular probes or generating nucleic acid fragments forsubsequent subcloning are known. Examples of techniques sufficient todirect persons of skill through such in vitro amplification methods,including the polymerase chain reaction (PCR) the ligase chain reaction(LCR), Qβ-replicase amplification and other RNA polymerase mediatedtechniques (e.g., NASBA) are found in Berger, Sambrook et al. (1989)Molecular Cloning--A Laboratory Manual (2nd Ed) Vol. 1-3; and Ausubel,as well as Mullis et al., (1987) U.S. Pat. No. 4,683,202; PCR ProtocolsA Guide to Methods and Applications (Innis et al. eds) Academic PressInc. San Diego, Calif. (1990) (Innis); Arnheim & Levinson (Oct. 1, 1990)C&EN 36-47; The Journal Of NIH Research (1991) 3, 81-94; (Kwoh et al.(1989) Proc. Natl. Acad. Sci. USA 86, 1173; Guatelli et al. (1990) Proc.Natl. Acad. Sci. USA 87, 1874; Lomell et al. (1989) J. Clin. Chem 35,1826; Landegren et al., (1988) Science 241, 1077-1080; Van Brunt (1990)Biotechnology 8, 291-294; Wu and Wallace, (1989) Gene 4, 560; Barringeret al. (1990) Gene 89, 117, and Sooknanan and Malek (1995) Biotechnology13: 563-564. Improved methods of cloning in vitro amplified nucleicacids are described in Wallace et al., U.S. Pat. No. 5,426,039.

Oligonucleotides for use as probes, e.g., in in vitro rOnc nucleic acidamplification methods, or for use as nucleic acid probes to detect rOncnucleic acids are typically synthesized chemically according to thesolid phase phosphoramidite triester method described by Beaucage andCaruthers (1981), Tetrahedron Letts., 22(20):1859-1862, e.g., using anautomated synthesizer, e.g., as described in Needham-VanDevanter et al.(1984) Nucleic Acids Res., 12:6159-6168. Oligonucleotides can also becustom made and ordered from a variety of commercial sources known topersons of skill. Purification of oligonucleotides, where necessary, istypically performed by either native acrylamide gel electrophoresis orby anion-exchange HPLC as described in Pearson and Regnier (1983) J.Chrom. 255:137-149. The sequence of the synthetic oligonucleotides canbe verified using the chemical degradation method of Maxam and Gilbert(1980) in Grossman and Moldave (eds.) Academic Press, New York, Methodsin Enzymology 65:499-560.

One of skill will recognize many ways of generating desired alterationsin a given nucleic acid sequence. Such well-known methods includesite-directed mutagenesis, PCR amplification using degenerateoligonucleotides, exposure of cells containing the nucleic acid tomutagenic agents or radiation, chemical synthesis of a desiredoligonucleotide (e.g., in conjunction with ligation and/or cloning togenerate large nucleic acids) and other well-known techniques. See,Giliman and Smith (1979) Gene 8:81-97; Roberts et al. (1987) Nature328:731-734 and Sambrook et al. (1989) Molecular Cloning--A LaboratoryManual (2nd Ed) Vol. 1-3; Innis, Ausbel, Berger, Needham VanDevanter andMullis (all supra).

Polypeptides of the invention can be synthetically prepared in a widevariety of well-known ways. Polypeptides of relatively short size aretypically synthesized in solution or on a solid support in accordancewith conventional techniques. See, e.g., Merrifield (1963) J. Am. Chem.Soc. 85:2149-2154. Various automatic synthesizers and sequencers arecommercially available and can be used in accordance with knownprotocols. See, e.g., Stewart and Young (1984) Solid Phase PeptideSynthesis, 2d. ed., Pierce Chemical Co. Polypeptides are also producedby recombinant expression of a nucleic acid encoding the polypeptidefollowed by purification using standard techniques.

Making Conservative Modifications of the Nucleic Acids and Polypeptidesof the Invention

One of skill will appreciate that many conservative variations of thesequences disclosed yield a substantially identical rOnc. For example,due to the degeneracy of the genetic code, "silent substitutions" (i.e.,substitutions of a nucleic acid sequence which do not result in analteration in an encoded polypeptide) are an implied feature of everynucleic acid sequence which encodes an amino acid. Similarly,conservative amino acid substitutions, in one or a few amino acids in anamino acid sequence are substituted with different amino acids withhighly similar properties (see, the definitions section, supra), arealso readily identified as being highly similar to a disclosed aminoacid sequence, or to a disclosed nucleic acid sequence which encodes anamino acid. Such conservatively substituted (or modified) variations ofeach explicitly disclosed sequence are a feature of the presentinvention.

One of skill will recognize many ways of generating alterations in agiven nucleic acid sequence. Such well-known methods includesite-directed mutagenesis, PCR amplification using degenerateoligonucleotides, exposure of cells containing the nucleic acid tomutagenic agents or radiation, chemical synthesis of a desiredoligonucleotide (e.g., in conjunction with ligation and/or cloning togenerate large nucleic acids) and other well-known techniques. See,Giliman and Smith (1979) Gene 8:81-97, Roberts et al. (1987) Nature328:731-734 and Sambrook, Innis, Ausbel, Berger, Needham VanDevanter andMullis (all supra).

Most commonly, polypeptide sequences are altered by changing thecorresponding nucleic acid sequence and expressing the polypeptide.However, polypeptide sequences are also optionally generatedsynthetically using commercially available peptide synthesizers toproduce any desired polypeptide (see, Merrifield, and Stewart and Young,supra).

One of skill can select a desired nucleic acid or polypeptide of theinvention based upon the sequences provided and upon knowledge in theart regarding ribonucleases generally. The physical characteristics andgeneral properties of RNases are known to skilled practitioners. Thespecific effects of some mutations in RNases are known. Moreover,general knowledge regarding the nature of proteins and nucleic acidsallows one of skill to select appropriate sequences with activitysimilar or equivalent to the nucleic acids and polypeptides disclosed inthe sequence listings herein. The definitions section herein describesexemplary conservative amino acid substitutions.

Finally, most modifications to nucleic acids and polypeptides areevaluated by routine screening techniques in suitable assays for thedesired characteristic. For instance, changes in the immunologicalcharacter of a polypeptide can be detected by an appropriateimmunological assay. Modifications of other properties such as nucleicacid hybridization to a target nucleic acid, redox or thermal stabilityof a protein, thermal histeresis, hydrophobicity, susceptibility toproteolysis, or the tendency to aggregate are all assayed according tostandard techniques.

rOnc Fusion Proteins and Other Therapeutic Moieties

The rOnc molecules may also include pharmacological agents orencapsulation systems containing various pharmacological agents. Theytypically will include a ligand to act as a targeting molecule to directthe rOnc to desired cells. The rOnc may be attached directly to a ligandor an antisense molecule which will assist in delivering the rOnc. See,for example, SEQ ID NOS:40-61. The rOnc can also be engineered tocontain a nuclear localization signal ("NLS") such as that described inamino acid positions 1 to 7 in SEQ ID NO:32 (and SEQ ID NO:31) to directthe rOnc within the cell. Alternatively, the Met at position 8 and thecorresponding nucleic acids at positions 22-24 of SEQ ID NO:31 has beenand can be omitted. The nucleic acid sequence for the NLS is nucleicacids 1-21 of SEQ ID NO:31. A signal peptide is also exemplified atamino acid positions 1-25 of SEQ ID NO:63.

The rOnc molecules are uniquely adapted for gene therapy applications.They can be fused to other therapeutic agents, for example, they couldbe fused to an anti-B cell lymphoma antibody. For example, as will beexplained in more detail below, rOnc molecules recombinantly fused to ananti-transferrin receptor antibody or an anti-colon cancer antibody wereactive. As mentioned above, nOnc has anti-tumor effects in vivo andpreferentially kills rapidly dividing cells stimulated by serum orgrowth promoting agents such as ras. The molecules are readilyinternalized in the cell. Their activity can be further facilitated byjoining them to a nuclear localization signal and the like to redirectthe molecules within the cell. Of particular use in tumor cells would beto target telomerase, an enzyme subject to degradation by RNase.

We have found that Onc synergizes with ras in microinjection studies.This means that Onc and ras have to be together in the cell. Onc doesnot synergize with ras when it enters the cell via its own routing. ACAAX (SEQ ID NO:33) motif is required to localize ras at the plasmamembrane (C=Cys, A=an aliphatic amino acid, X=S,M,C,A, or Q, an exampleis Cys-Val-Ile-Met (SEQ ID NO:34)). Importantly this type of sequencehas been shown to target heterologous proteins to the plasma membrane(Hancock, J., Cadwallader, K., Paterson, H. and C. Marshall (1991) EMBOJ. 10:4033). It would be desirable to join the rOnc gene with DNAencoding a CAAX (SEQ ID NO:33) signal as given in the example, or KDELas described below.

Telomerase is being investigated as a "universal cancer target" (G. B.Morin, JNCI. (1995) 87:859). It is an RNA protein that is located in thenucleus. It has been shown that antisense to telomerase RNA can inhibitthe function of the enzyme and block the growth of cancer cells (J. Fenget al., Science (1995) 269:1236). RNase can also destroy the activity ofthe enzyme. Onc can also destroy the activity of the enzyme whenincubated with a cell extract containing telomerase. An NLS/Onc molecule(such as that set out in SEQ ID NO: 32) can be made to route Onc to thenucleus so that it can degrade telomerase. The NLS we used has beenshown to redirect proteins to the nucleus for the aim of interferingwith the function of a nuclear antigen (S. Biocca, M. S. Neuberger andA. Cattaneo, (1990) 9:101). Our NLS/Onc molecule is effective in killingcells.

An amino terminal sequence to the recombinant molecule may be preferredwhere it is desirable to translocate the molecule into the cytosol oftarget cells. Such signal peptide is typically inserted at the amino endof the protein. For example, the first amino acids of the recombinantmolecules described herein (after Met) could be KDEL (SEQ ID NO:64) andwould accomplish signalling the molecule to the endoplasmic reticulum.Amino acid sequences which include KDEL, repeats of KDEL, or othersequences that function to maintain or recycle proteins into theendoplasmic reticulum, referred to here as "endoplasmic retentionsequences" may be employed.

Optionally, the rOnc molecule attached to a ligand may include anencapsulation system, such as a liposome or micelle that contains anadditional therapeutic composition such as a drug, a nucleic acid (e.g.an antisense nucleic acid), or another therapeutic moiety that ispreferably shielded from direct exposure to the circulatory system.Means of preparing liposomes attached to antibodies are well known tothose of skill in the art. See, for example, U.S. Pat. No. 4,957,735,Connor et al., Pharm. Ther., 28: 341-365 (1985).

One of skill will appreciate that the ligand molecule or othertherapeutic component and the rOnc molecule may be joined together inany order. Thus, where the ligand is a polypeptide, the rOnc moleculemay be joined to either the amino or carboxy termini of the ligand ormay also be joined to an internal region of either molecule as long asthe attachment does not interfere with the respective activities of themolecules.

The molecules may be attached by any of a number of means well-known tothose of skill in the art. Typically the rOnc will be conjugated, eitherdirectly or through a linker (spacer), to the ligand. However, whereboth the rOnc and the ligand or other therapeutic are polypeptides it ispreferable to recombinantly express the chimeric molecule as asingle-chain fusion protein.

In one embodiment, the rOnc molecule is chemically conjugated to anothermolecule (e.g. a cytotoxin, a label, a ligand, or a drug or liposome).Means of chemically conjugating molecules are well-known to those ofskill.

The procedure for attaching an agent to an antibody or other polypeptidetargeting molecule will vary according to the chemical structure of theagent. Polypeptides typically contain a variety of functional groups;e.g., carboxylic acid (COOH) or free amine (--NH₂) groups, which areavailable for reaction with a suitable functional group on an rOncmolecule to bind the other molecule thereto.

Alternatively, the ligand and/or rOnc molecule may be derivatized toexpose or attach additional reactive functional groups. Thederivatization may involve attachment of any of a number of linkermolecules such as those available from Pierce Chemical Company, RockfordIll.

A "linker", as used herein, is a molecule that is used to join twomolecules. The linker is capable of forming covalent bonds to bothmolecules. Suitable linkers are well known to those of skill in the artand include, but are not limited to, straight or branched-chain carbonlinkers, heterocyclic carbon linkers, or peptide linkers. Where bothmolecules are polypeptides, the linkers may be joined to the constituentamino acids through their side groups (e.g., through a disulfide linkageto cysteine). However, in a preferred embodiment, the linkers will bejoined to the alpha carbon amino and carboxyl groups of the terminalamino acids.

A bifunctional linker having one functional group reactive with a groupon a particular agent, and another group reactive with an antibody, maybe used to form a desired immunoconjugate. Alternatively, derivatizationmay involve chemical treatment of the ligand, e.g., glycol cleavage ofthe sugar moiety of a glycoprotein antibody with periodate to generatefree aldehyde groups. The free aldehyde groups on the antibody may bereacted with free amine or hydrazine groups on an agent to bind theagent thereto. (See U.S. Pat. No. 4,671,958). Procedures for generationof free sulfhydryl groups on polypeptides, such as antibodies orantibody fragments, are also known (See U.S. Pat. No. 4,659,839).

Many procedure and linker molecules for attachment of various compoundsincluding radionuclide metal chelates, toxins and drugs to proteins suchas antibodies are known. See, for example, European Patent ApplicationNo. 188,256; U.S. Pat. Nos. 4,671,958, 4,659,839, 4,414,148, 4,699,784;4,680,338; 4,569,789; and 4,589,071; and Borlinghaus et al. Cancer Res.47: 4071-4075 (1987) which are incorporated herein by reference. Inparticular, production of various immunotoxins is well-known within theart and can be found, for example in "Monoclonal Antibody-ToxinConjugates: Aiming the Magic Bullet," Thorpe et al., MonoclonalAntibodies in Clinical Medicine, Academic Press, pp. 168-190 (1982),Waldmann, Science, 252: 1657 (1991), U.S. Pat. Nos. 4,545,985 and4,894,443 which are incorporated herein by reference.

In some circumstances, it is desirable to free the rOnc from the ligandwhen the chimeric molecule has reached its target site. Therefore,chimeric conjugates comprising linkages which are cleavable in thevicinity of the target site may be used when the effector is to bereleased at the target site. Cleaving of the linkage to release theagent from the ligand may be prompted by enzymatic activity orconditions to which the immunoconjugate is subjected either inside thetarget cell or in the vicinity of the target site. When the target siteis a tumor, a linker which is cleavable under conditions present at thetumor site (e.g. when exposed to tumor-associated enzymes or acidic pH)may be used.

A number of different cleavable linkers are known to those of skill inthe art. See U.S. Pat. Nos. 4,618,492; 4,542,225, and 4,625,014. Themechanisms for release of an agent from these linker groups include, forexample, irradiation of a photolabile bond and acid-catalyzedhydrolysis. U.S. Pat. No. 4,671,958, for example, includes a descriptionof immunoconjugates comprising linkers which are cleaved at the targetsite in vivo by the proteolytic enzymes of the patient's complementsystem. In view of the large number of methods that have been reportedfor attaching a variety of radiodiagnostic compounds, radiotherapeuticcompounds, drugs, toxins, and other agents to antibodies one skilled inthe art will be able to determine a suitable method for attaching agiven agent to an antibody or other polypeptide.

Production of rOnc Molecules or Fusion Proteins

Where the molecules of interest are relatively short (i.e., less thanabout 50 amino acids) they may be synthesized using standard chemicalpeptide synthesis techniques. Where two molecules of interest arerelatively short the chimeric molecule may be synthesized as a singlecontiguous polypeptide. Alternatively the molecules may be synthesizedseparately and then fused by condensation of the amino terminus of onemolecule with the carboxyl terminus of the other molecule therebyforming a peptide bond. Alternatively, the molecules may each becondensed with one end of a peptide spacer molecule thereby forming acontiguous fusion protein.

Solid phase synthesis in which the C-terminal amino acid of the sequenceis attached to an insoluble support followed by sequential addition ofthe remaining amino acids in the sequence is the preferred method forthe chemical synthesis of the polypeptides of this invention. Techniquesfor solid phase synthesis are described by Barany and Merrifield,Solid-Phase Peptide Synthesis; pp. 3-284 in The Peptides: Analysis,Synthesis, Biology. Vol. 2: Special Methods in Peptide Synthesis, PartA., Merrifield, et al. J. Am. Chem. Soc., 85: 2149-2156 (1963), andStewart et al., Solid Phase Peptide Synthesis, 2nd ed. Pierce Chem. Co.,Rockford, Ill. (1984) which are incorporated herein by reference.

In a preferred embodiment, the chimeric fusion proteins of the presentinvention are synthesized using recombinant DNA methodology. Generallythis involves creating a DNA sequence that encodes the fusion protein,placing the DNA in an expression cassette under the control of aparticular promoter, expressing the protein in a host, isolating theexpressed protein and, if required, renaturing the protein.

DNA encoding the fusion proteins of this invention, as well as the rOncmolecules themselves, may be prepared by any suitable method, including,for example, cloning and restriction of appropriate sequences or directchemical synthesis by methods such as the phosphotriester method ofNarang et al. Meth. Enzymol. 68: 90-99 (1979); the phosphodiester methodof Brown et al., Meth. Enzymol. 68: 109-151 (1979); thediethylphosphoramidite method of Beaucage et al., Tetra. Lett., 22:1859-1862 (1981); and the solid support method of U.S. Pat. No.4,458,066, all incorporated by reference herein.

Chemical synthesis produces a single-stranded oligonucleotide. This maybe converted into double-stranded DNA by hybridization with acomplementary sequence, or by polymerization with a DNA polymerase usingthe single strand as a template. One of skill would recognize that whilechemical synthesis of DNA is limited to sequences of about 100 bases,longer sequences may be obtained by the ligation of shorter sequences.

Alternatively, subsequences may be cloned and the appropriatesubsequences cleaved using appropriate restriction enzymes. Thefragments may then be ligated to produce the desired DNA sequence.

In a preferred embodiment, DNA encoding fusion proteins or rOncmolecules of the present invention may be cloned using DNA amplificationmethods such as polymerase chain reaction (PCR). If two molecules arejoined together, one of skill will appreciate that the molecules may beseparated by a peptide spacer consisting of one or more amino acids.Generally the spacer will have no specific biological activity otherthan to join the proteins or to preserve some minimum distance or otherspatial relationship between them. However, the constituent amino acidsof the spacer may be selected to influence some property of the moleculesuch as the folding, net charge, or hydrophobicity.

The nucleic acid sequences encoding the rOnc molecules or the fusionproteins may be expressed in a variety of host cells, including E. coli,other bacterial hosts, yeast, and various higher eukaryotic cells suchas the COS, CHO and HeLa cells lines and myeloma cell lines. Therecombinant protein gene will be operably linked to appropriateexpression control sequences for each host. For E. coli this includes apromoter such as the T7, trp, or lambda promoters, a ribosome bindingsite and preferably a transcription termination signal. For eukaryoticcells, the control sequences will include a promoter and preferably anenhancer derived from immunoglobulin genes, SV40, cytomegalovirus, etc.,and a polyadenylation sequence, and may include splice donor andacceptor sequences.

The expression vectors or plasmids of the invention can be transferredinto the chosen host cell by well-known methods such as calcium chloridetransformation for E. coli and calcium phosphate treatment orelectroporation for mammalian cells. Cells transformed by the plasmidscan be selected by resistance to antibiotics conferred by genescontained on the plasmids, such as the amp, gpt, neo and hyg genes.

Once expressed, the recombinant rOnc or fusion proteins can be purifiedaccording to standard procedures of the art, including ammonium sulfateprecipitation, affinity columns, column chromatography, gelelectrophoresis and the like (see, generally, R. Scopes, ProteinPurification, Springer-Verlag, N.Y. (1982), Deutscher, Methods inEnzymology Vol. 182: Guide to Protein Purification., Academic Press,Inc. N.Y. (1990)). Substantially pure compositions of at least about 90to 95% homogeneity are preferred, and 98 to 99% or more homogeneity aremost preferred for pharmaceutical uses. Once purified, partially or tohomogeneity as desired, the polypeptides may then be usedtherapeutically.

Accordingly, this invention also provides for host cells and expressionvectors comprising the nucleic acid sequences described above.

Further, the present invention includes a method of selectively killingcells using a rOnc joined to a ligand to create a selective cytotoxicreagent of the present invention. The method comprises contacting thecells to be killed with a cytotoxic reagent of the present inventionhaving a ligand binding moiety that specifically delivers the reagent tothe cells to be killed. This method of the present invention may be usedfor cell separation in vitro by selectively killing unwanted types ofcells, for example, in bone marrow prior to transplantation into apatient undergoing marrow ablation by radiation, for killing leukemiacells or T-cells that would cause graft-versus-host disease.

For methods of use in vivo, preferably the mammalian protein of thereagent used in this method is endogenous to the species in which thereagent is intended for use. Preferably, for use in humans, thecytotoxic reagent is a fusion protein comprising a humanized chimericantibody and a humanized rOnc. Specific in vivo methods of thisinvention include a method for the chemotherapeutic alleviation ofcancer in mammals comprising administering a cytotoxic amount of aselective cytotoxic reagent according to the present invention. Themethods are particularly useful for treating tumors sensitive to thecytotoxic reagents. Tumors of particular interest include pancreatic,colon, breast and kidney tumors.

Pharmaceutical Compositions

The rOnc molecules and fusion proteins employing them of this inventionare useful for parenteral, topical, oral, or local administration, suchas by aerosol or transdermally, for prophylactic and/or therapeutictreatment. The pharmaceutical compositions can be administered in avariety of unit dosage forms depending upon the method ofadministration. For example, unit dosage forms suitable for oraladministration include powder, tablets, pills, capsules and lozenges. Itis recognized that the subject molecules and fusion proteins andpharmaceutical compositions of this invention, when administered orally,must be protected from digestion. This is typically accomplished eitherby complexing the protein with a composition to render it resistant toacidic and enzymatic hydrolysis or by packaging the protein in anappropriately resistant carrier such as a liposome. Means of protectingproteins from digestion are well known in the art.

The pharmaceutical compositions of this invention are particularlyuseful for parenteral administration, such as intravenous administrationor administration into a body cavity or lumen of an organ. Thecompositions for administration will commonly comprise a solution of thechimeric molecule dissolved in a pharmaceutically acceptable carrier,preferably an aqueous carrier. A variety of aqueous carriers can beused, e.g., buffered saline and the like. These solutions are sterileand generally free of undesirable matter. These compositions may besterilized by conventional, well known sterilization techniques. Thecompositions may contain pharmaceutically acceptable auxiliarysubstances as required to approximate physiological conditions such aspH adjusting and buffering agents, toxicity adjusting agents and thelike, for example, sodium acetate, sodium chloride, potassium chloride,calcium chloride, sodium lactate and the like. The concentration oftherapeutic molecule in these formulations can vary widely, and will beselected primarily based on fluid volumes, viscosities, body weight andthe like in accordance with the particular mode of administrationselected and the patient's needs.

Thus, a typical pharmaceutical composition for intravenousadministration would be about 0.1 to 10 mg per patient per day. Dosagesfrom 0.1 up to about 100 mg per patient per day may be used,particularly when the drug is administered to a secluded site and notinto the blood stream, such as into a body cavity or into a lumen of anorgan. Actual methods for preparing parenterally administrablecompositions will be known or apparent to those skilled in the art andare described in more detail in such publications as Remington'sPharmaceutical Science, 15th ed., Mack Publishing Company, Easton, Pa.(1980).

The compositions containing the present rOnc molecules or the fusionproteins or a cocktail thereof (i.e., with other proteins) can beadministered for therapeutic treatments. In therapeutic applications,compositions are administered to a patient suffering from a disease, ina cytotoxic amount, an amount sufficient to kill cells of interest. Anamount adequate to accomplish this is defined as a "therapeuticallyeffective dose." Amounts effective for this use will depend upon theseverity of the disease and the general state of the patient's health.

Single or multiple administrations of the compositions may beadministered depending on the dosage and frequency as required andtolerated by the patient. In any event, the composition should provide asufficient quantity of the proteins of this invention to effectivelytreat the patient.

All patents, applications and publications cited herein are incorporatedby reference herein. The following examples are provided forillustrative purposes only and are not to be construed as limiting theinvention in any way.

EXAMPLES Example I Cloning and Expression of rOnc and Onc Conjugateswith EDN

A. Materials. Native ONCONASE® ("nOnc") (SEQ ID NO:1) Ardelt et al.(1991) J. Biol. Chem. 256, 245-251 and recombinant human EDN ("rEDN")(SEQ ID NO:9) Newton et al. (1994) J Biol Chem. 269, 26739-26745 werepurified from Rana pipiens oocytes, NASCO, Fort Atkinson, Wis. andEscherichia coli, respectively, as described. Antibodies to thedenatured proteins were prepared by Assay Research, Inc., College Park,Md. Reagents for performing PCR, and direct cloning of PCR products,were obtained from Perkin-Elmer Corp., Norwalk, Conn. and fromInvitrogen, San Diego, Calif. respectively. Substrates for theribonuclease assays were purchased from Sigma, St. Louis, Mo. andBoehringer Mannheim, Indianapolis, Ind. The materials and their sourcesused in the construction and expression of the recombinant proteins aswell as the rabbit reticulocyte lysate are described by Newton et al.,Biochemistry 35:545 (1996).

B. PCR Cloning of Onconase. Rana pipiens genomic DNA was isolatedaccording to standard procedures using proteinase K Maniatis, T.,Fritsch, E. F. & Sambrook, J. Molecular Cloning, a laboratory manual(Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1982). Aseries of degenerate primers were designed to correspond to amino acidsin various regions of the published nOnc sequence Ardelt et al. (1991)J. Biol. Chem. 256, 245-251. The PCR reaction was performed according tothe manufacturer's instructions using 15 μg of genomic DNA in 100 μl.All reagents except the DNA were combined and incubated at 95° C. for 8min to inactivate any residual proteinase K before the addition of theTaq DNA polymerase. PCR was performed for 40 cycles of denaturation at94° C. for 1 min, annealing for 2 min at 55° C. and primer extension for2 min at 72° C. Several pairs of primers yielded products of theexpected size. The largest product (252 bp) was obtained using theforward primer encoding amino acid residues 15-23(AG(GA)GATGT(GT)GATTG(TC)GATAA(CT)ATCATG) (SEQ ID NO:35) and the reverseprimer encoding amino acid residues 90-98(TGTGA(AG)AA(CT)CAGGC(AC)CC(TA)GT(GT)CA(CT)TTT) (SEQ ID NO:36). Thisfragment was subcloned into pCR™II by TA cloning and a clone carrying aninsert of the appropriate size was directly sequenced and found toencode amino acid residues 16-98 of nOnc ("Rana 9") (SEQ ID NO:2). Thecorresponding nucleic acid sequence is set out in SEQ ID NO:37.

C. Plasmid Construction, Expression, Protein Purification and in VitroAssays. The N- and C-termini of nOnc were reconstructed using the PCRtechnique of splicing by overlap extension Horten et al. (1990)BioTechniques 8, 528-532 with amino acid residues 1-15 of nOnc or aminoacid residues 1-21 of EDN at the N-terminal and amino acid residues99-104 of nOnc at the C-terminal. The assembled genes were insertedbetween the XbaI and BamHI sites of the bacterial expression vector,pET-11d, Novagen, Madison, Wis. All procedures were accomplishedessentially as described in Newton et al. (1994) J Biol Chem. 269,26739-26745. The plasmids were expressed in BL21(DE3) E. coli cells asrecommended by the supplier, Novagen, Madison Wis. The fusion proteinswere isolated from inclusion bodies, denatured, renatured and dialyzedas described Newton et al. (1994) J Biol Chem. 269, 26739-26745 beforebeing applied to a CM-Sephadex C-50 column, Pharmacia Biotech Inc.,Piscataway, N.J. The proteins were eluted with a NaCl gradient (0-0.5M)in 20 mM Tris-HCl, pH 7.5, containing 10% glycerol. Final purificationto >95% was achieved by size exclusion chromatography on Sephadex G-100equilibrated and eluted with 5% formic acid. The proteins were pooled,concentrated by amicon ultrafiltration using a YM3 membrane (orlyophilized), Amicon, Beverly, Mass. and dialyzed against 20 mMTris-HCl, pH 7.5, containing 10% glycerol before being assayed.

Ribonuclease activity using high molecular weight RNA and tRNA wasdetermined following published protocols, Newton et al. (1994) JNeurosci 14, 538-544 at 37° C. through the formation of perchloric acidsoluble nucleotides following published protocols (Newton et al. (1996)Biochem. 35:545-553). With poly (A,C), UpG and poly U, ribonucleaseactivity was assayed spectrophotometrically according to DePrisco etal., and Libonati and Florida DePrisco et al. (1984) Biochimica etBiophysica Acta 788, 356-363, Libonati, M. & Floridi, A. (1969) EuropeanJ. Biochem. 8, 81-87. Briefly, activity was assayed by measuring theincrease in absorbance at 260 nm. Incubation mixtures (1 ml of 10 mMimidazole, 0.1 M NaCl, pH 6.5 or pH 7) contained substrate andappropriate amounts of enzyme solution at 25° C. The in vitrotranslation assay, St. Clair et al. (1987) Proc Natl Acad Sci 84,8330-8334, and the cell viability assays, Pearson et al. (1991) J NatlCancer Inst 83, 1386-1391, using the(3-[4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide; Thiazolylblue] (MTT) Mossman, T. (1983) J. Immunol. Methods 65, 55-63 wereperformed as previously described.

D. Cloning and Expression of [Met-(-1)] rOnc and rOnc chimeras. Eightdifferent oligonucleotide primers were designed to correspond tospecific regions in the primary amino acid structure of nOnc, Ardelt etal. (1991) J. Biol. Chem. 256, 245-251 and amplification of Rana pipiensgenomic DNA was carried out in a thermal cycler, as described above. Aprimer pair corresponding to amino acid residues 15 to 23 and 90 to 98of nOnc, respectively, generated a 252 bp fragment. That PCR product,here denoted Rana clone 9, was cloned into pCR™II and sequence analysisconfirmed that the PCR product encoded Onc (104 amino acids, total) fromamino acid residue 16 to 98 (FIG. 1).

The entire recombinant Onc ("rOnc") gene (SEQ ID NO:38) was constructedby PCR extension and cloned into an expression vector using methodologypreviously described Newton et al. (1994) J Biol Chem. 269, 26739-26745.The amino and carboxyl termini of rOnc were completed by inserting thefirst 15 and last 6 amino acid residues of nOnc, respectively. Theconfiguration of the semi-synthetic rOnc gene is depicted at the top ofFIG. 2A. The primers were designed to overlap with the DNA sequence ofthe Rana clone 9 PCR product. The plasmid was expressed in BL21(DE3) E.coli and the recombinant protein was isolated from inclusion bodies asdescribed in Newton et al. (1994) J Biol Chem. 269, 26739-26745 beforebeing applied to a CM Sephadex C-50 column. Final purification to >95%was achieved by size exclusion chromatography. The rOnc obtained fromthe bacteria in this expression system contains an extra methionine atthe amino terminal [Met-(-1)] (SEQ ID NO:39) in contrast to theauthentic pyroglutamyl amino acid residue (<Glu-1) of the native protein(SEQ ID NO:1).

To humanize [Met-(-1)] rOnc while maintaining the alignment of theactive site residues (FIG. 2B), the N-terminal of Rana clone 9 was alsoreconstituted with oligonucleotides that coded for the first 21 aminoacid residues of a human eosinophil RNase, EDN (FIG. 2B, rEDN.sub.(1-21)Onc). PCR cloning can result in sequence errors. Indeed, the DNAsequence of the gene encoding EDN.sub.(1-21) Onc contained an A to Gsubstitution resulting in a change from Asp to Gly at position 26 in thechimera (residue 20 in nOnc) and is designated as rEDN.sub.(1-21)rOncG₂₆ in FIG. 2B. Another plasmid containing encoding rEDN.sub.(1-21)rOnc was sequenced and found to have the correct DNA sequence. Since themutation resulted in the substitution of a charged amino acid with asmall neutral residue the mutant chimera was also expressed andcharacterized for activity. In addition, [Met-(-1)]rOnc was mutated atposition 20 from Asp to Gly (rOncGly₂₀, FIGS. 2A and 2B).

E. Ribonuclease activity of Onc, EDN, [Met-(-1)]rOnc and hybrid rOncproteins. Both nOnc (Lin, J. J., et al. (1994) Biochem Biophys ResCommun 204, 156-162) and EDN (Saxena et al. (1992) J. Biol. Chem. 267,21982-21986) are potent inhibitors of in vitro translation in the rabbitreticulocyte lysate by mechanisms that depend upon their respectivenucleolytic activities. As depicted in Table 1, the addition of nOnc orEDN to a rabbit reticulocyte lysate caused the inhibition of proteinsynthesis as measured by the incorporation of [³⁵ S]methionine into acidprecipitable protein. Whereas both nOnc and EDN inhibited proteinsynthesis with IC_(50s) of 0.2 and 1.3 ng/ml, respectively,[Met-(-1)]rOnc, [Met-(-1)]rOncG20, and rEDN.sub.(1-21) rOncG26 wereconsiderably less potent (IC_(50s) 98, 28 and 28 ng/ml, respectively).The least active RNase in this assay was rEDN.sub.(1-21) rOnc with anIC₅₀ of 1600 ng/ml. Placental ribonuclease inhibitor (PRI) binds tightlyto EDN and inhibits its enzymatic activity, Sorrentino et al. (1992) J.Biol. Chem 267, 14859-14865, yet nOnc activity is very little affectedby PRI, Wu, Y. N., et al. (1993) Journal of Biological Chemistry 268,10686-10693 and Table 1, despite its homology to EDN and other membersof the pancreatic RNase superfamily. In this regard, it is interestingthat the activity of rEDN.sub.(1-21) rOnc is, like nOnc, barely affectedby PRI while the hybrid RNase with the Gly mutation now behaves morelike EDN in that its activity is significantly inhibited (21 fold) byPRI.

The ribonuclease activity of these proteins was also assessed in assaysusing high and low molecular weight substrates. As shown in Table 2, EDNand nOnc have different substrate specificities consistent withpreviously published results (Ardelt et al. (1991) J. Biol. Chem. 256,245-251, Sorrentino et al. (1992) J. Biol. Chem 267, 14859-14865, Ardeltet al. (1994) Protein Sci 3, Suppl. 1, 137). Consistent with the resultspresented in Table 1, [Met-(-1)]rOnc (SEQ ID NO:39) and rEDN.sub.(1-21)rOnc were much less active with all of the substrates (non detectable orvery little activity under the assay conditions employed). Surprisingly,the Gly containing hybrid protein, manifested significant ribonucleaseactivity especially under conditions optimal for EDN enzymatic activity.EDN is more active at a neutral pH (Sorrentino et al. (1992) J. Biol.Chem 267, 14859-14865) and as seen in Table 2 there is a marked increasein EDN degradation of tRNA at pH 7.5 compared to pH 6 (42.3 fold). Also,behaving like EDN, the Gly-containing hybrid increases in activity witha pH shift from 6 to 7.5 (21.7 fold) while nOnc loses activity at pH 7.5consistent with its pH optimum that ranges from 6-6.5 (Ardelt et al.(1991) J. Biol. Chem. 256, 245-251, Ardelt et al. (1994) Protein Sci 3,Suppl. 1, 137). The enhanced EDN-like activity of the Gly-containinghybrid protein is also evidenced by its behavior with poly(A,C) which isan excellent substrate for EDN. As seen in Table 2, only rEDN.sub.(1-21)rOncG26 expresses almost 50% of the enzymatic activity of EDN with thissubstrate whereas the activity of the other RNases are negligible.Similar results were observed with poly(U). In contrast, there was nodetectable activity of rEDN or rEDN.sub.(1-21) rOncG26 with UpG, anoptimal Onconase substrate (Ardelt et al. (1994) Protein Sci 3, Suppl.1, 137). In summary, both [Met-(-1)]rOnc and rEDN.sub.(1-21) rOnc areless enzymatically active than nOnc or rEDN. Although, rEDN.sub.(1-21)rOncG26 expresses significant EDN-like enzymatic activity when assayedusing defined substrates and conditions optimal for EDN, it is not asactive as EDN in any assay. This could result from an impaired enzymesubstrate interaction or from the use of suboptimal assay conditions forthis hybrid enzyme.

                  TABLE 1                                                         ______________________________________                                        Activity of [Met-(-1)]rOnc or Hybrid Proteins in Rabbit Reticulocyte          Lysate compared to rEDN or nOnc in the Presence or Absence of PRI                        IC.sub.50.sup.a (ng/ml)                                                       (-)PRI  (+)PRI  Fold Difference                                    ______________________________________                                        nOnc         0.2       0.24    1.2                                            rEDN         1.3       >40     >30.7                                          [Met-(-1)]rOnc                                                                             96        140     1.4                                            [Met-(-1)]rOncG20                                                                          28        24      0.9                                            rEDN.sub.(1-21) rOnc                                                                       1600      3200    2                                              rEDN.sub.(1-21) rOncG26                                                                    28        600     21                                             ______________________________________                                         .sup.a IC.sub.50 is the concentration of protein necessary to inhibit         protein synthesis by 50% in the rabbit reticulocyte lysate. Data points       result from the average of at least three assays.                        

                                      TABLE 2                                     __________________________________________________________________________    Activity of RNases on Different Substrates                                                        RNase Activity (units/mg protein).sup.a                   Substrate                                                                           Assay pH                                                                           rEDN nOnc                                                                              [Met-(-1)]rOnc                                                                       rEND.sub.1-21r Onc                                                                   rEDN.sub.1-21r OncG.sub.26                  __________________________________________________________________________    Yeast RNA.sup.a                                                                     6.0  6000 560 0.01   8      120                                         tRNA.sup.a,c                                                                        6.0  1100 390 12     4      340                                         tRNA.sup.a,c                                                                        7.5  46000                                                                              60  50     130    7400                                        poly (A,C).sup.b                                                                    7.0  8000 0.04                                                                              5      4.5    3900                                        UpG.sup.b                                                                           6.5  0.05 0.18                                                                              <0.01  <0.01  <0.01                                       poly U.sup.b                                                                        7.0  16.5 0.15                                                                              0.20   0.35   4.5                                         __________________________________________________________________________     .sup.a RNase activity was quantitated through the formation of perchloric     acid soluble nucleotides. Units are defined as the changes in A.sub.260       per minute calculated from the slopes of the linear part of the assays.       Each value is the average of 2-3 assays in separate experiments.              .sup.b Spectrophotometric assays were performed according to Deprisco et      al. (1984) and Libonati and Floridi (1969) as described in Materials and      Methods. Units are defined as the changes in A.sub.260 per minute             calculated from the slopes of the linear part of the assays. Each value i     the average of two or more determinatinos.                                    .sup.c [Met(-1)]rOncG20 had no detectable activity.                      

F. Inhibition of protein synthesis in four human tumor cell lines byRNases. The cytotoxic effect of [Met-(-1)]rOnc and the two hybrid RNaseswere compared to rEDN and nOnc by determining cell viability using theMTT assay. As depicted in FIG. 3, nOnc decreased tumor cell viability inall four human tumor cell lines. At the concentrations shown, rEDN hadno effect on the viability of any of the cell lines. In contrast tonOnc, [Met-(-1)]rOnc as well as [Met-(-1)]rOncG20 was consistently lesscytotoxic in all four cell lines. Yet, rEDN.sub.(1-21) rOncG26 was morecytotoxic than nOnc in ACHN, human renal carcinoma cells and equallycytotoxic in the MDA-MB-231 human breast carcinoma cell line. AlthoughrEDN.sub.(1-21) rOncG26 was less active than nOnc in the SF-539 and HS578T human glioma and breast cancer cell lines, respectively, it wasstill more active than [Met-(-1)]rOnc or rEDN.sub.(1-21) rOnc proteincontaining Asn at position 26.

G. Structural Analysis of the hybrid RNases. Modeling the hybrid RNasewas based on the alignment of the structures for Onc (Mosimann S. C.,Ardelt W., James M. N. G., (1994), Refined 1.7 A X-ray crystallographicstructure of P-30 protein, an amphibian ribonuclease with anti-tumoractivity (J Mol Biol 236, 1141-1153) and EDN (Mosimann S. C., Newton D.L., Youle R. J., James M., X-ray crystallographic structure ofrecombinant eosinophil-derived neurotoxin at 1.83A resolution J MolBiol). This and subsequent alignments were carried out using ALIGN(Satow Y., Cohen G. H., Padlan E. A., Davies D. R., (1986), J. Mol Biol190, 593-604).

H. Modeling the structures of the hybrid RNases. The coordinates for Oncand EDN were superimposed on the basis of C.sup.α trace alignment.Residues in conserved zones, particularly in the active site, showedvery little displacement when comparing both structures (global r.m.s.d.of 1.44 A for 90 C.sup.α atom pairs). The hybrid protein was modeled bymanual rebuilding and geometry regularization using TOM (Cambillau C.,Horjales E., (1987), J. Mol Graph 5, 174-177). Subsequently, the modelsfor rEDN.sub.(1-21) rOnc and rEDN.sub.(1-21) rOncG26 were assigned anoverall B-factor of 15 A² for all non-hydrogen atoms and independentlysubject to 300 cycles of positional energy minimization with the programXPLOR (Brunger A. (1992) XPLOR: a system for X-ray crystallography andNMR., New Haven: Yale University Press). The minimization yieldedvirtually identical structures in both cases, the highest distance basedon C.sup.α trace alignment being 0.44 for the C.sup.α of the mutatedresidue 26. The geometry quality of the final models were assessed withPROCHECK (Laskowski R. A., MacArthur M. W., Moss D. S., Thornton J. M.,(1993), J Appl Crystallogr 26, 283-291).

The structural basis for the marked differences in activity between theGly and Asp containing hybrid RNases are not obvious from modeling theseproteins especially since residue 26 is distant from the active site.When the highly homologous structure of RNase A complexed with apentanucleotide (Fontecilla-Camps J. C., deLorens R., leDu M. H.,Cuchillo C. M., (1994), J. Biol Chem 269, 21526-21531) was superimposedon the structure of the hybrid protein model, the nucleotide wasobserved also to be distant from the region of the mutation. However,the arrangement of the polynucleotide chain in the different RNases doesnot necessarily have to coincide. In the structure of EDN, a secondsulfate ion was found in addition to the one in the active site(Mosimann S. C., Newton D. L., Youle R. J., James M., X-raycrystallographic structure of recombinant eosinophil-derived neurotoxinat 1.83A resolution J Mol Biol). This second sulfate is likely replacinga phosphate from the nucleotide to be cleaved, but no phosphate ion islocated in the equivalent position in the RNase A-pentanucleotidecomplex. Moreover, one of the phosphates in this complex is forming asalt bridge with Lys-66, a residue which has no counterpart in Onc sinceit is located in a loop with a different topology in both molecules.Thus, whether the difference in enzymatic activity between the Asp andGly mutants in the chimera is related to a change in the bindingaffinity for the substrate remains an open question.

Although the structural basis for the difference in the activities ofthe two EDN-Onc hybrids is not clear, the EDN-like behavior of therEDN.sub.(1-21) rOncG26 hybrid can likely be attributed to theconfiguration of the N-terminal region since both the pyroglutamic acidin nOnc and Lys-1 in EDN are located in the area of the active site(Mosimann S. C., Ardelt W., James M. N. G., (1994), Refined 1.7 A X-raycrystallographic structure of P-30 protein, an amphibian ribonucleasewith anti-tumor activity J Mol Biol 236, 1141-1153; Mosimann S. C.,Newton D. L., Youle R. J., James M., X-ray crystallographic structure ofrecombinant eosinophil-derived neurotoxin at 1.83A resolution J MolBiol). In addition, the introduction of a Gly mutation in [Met-(-1)]rOncdid not significantly affect enzymatic activity. The preference of Uover C in the B1 subsite of RNase A has been related to the presence ofa particular residue (Asp-83) (DelCardayre S. B., Raines R. T., (1995),A residue to residue hydrogen bond mediates the nucleotide specificityof ribonuclease A J Mol Biol 252, 328-336). The corresponding residue innOnc is also an aspartic acid (Asp-67), while in EDN this position isoccupied by a histidine (His-82). EDN is more active toward poly (A,C),suggesting that it "prefers" C in the B1 subsite, possibly because itcontains a histidine residue as opposed to the aspartic acid in nOnc andRNase A. Taken together, this could explain the decreased activity ofthe Gly containing hybrid relative to rEDN since, according to thishypothesis, the presence of the Asp residue contributed by the rOncsequence would favor the binding of U over C. With regard to thedifference in PRI inhibition, the superposition between the hybridproteins and RNase A demonstrates that Asp-26 in the EDN-Onc chimeras isin the equivalent position to Asn-27 in RNase A that has been reportedto be in contact with PRI (Kobe B., Deisenhofer J., (1995), Nature 374,183-186). In addition, Asp-24 in both chimeras is very close to thisregion. Thus, the accumulation of negative charges in this area couldprevent binding by the inhibitor. If so, the substitution of Gly for Aspwould decrease the negative charge and restore the binding capacity.

Example II rOnc-Antibody Fusion Proteins

Additional rOnc-antibody and ligand proteins have been produced and arehighly active. E6FB[Met-(-1)]SerrOnc is an rOnc molecule having theamino acid sequence set out in SEQ ID NO:64 and includes the Fv sequencefrom antibody E6, an anti-transferrin receptor antibody. See sequencesfor E6 at amino acid positions 1-237 in SEQ ID NO:41. "FB" refers to alinker used to link the antibody and the rOnc portion of the moleculeand is found at nucleic acid positions 712 through 750 in SEQ ID NO:40.E6FB[Met-(-1)]SerrOnc (SEQ ID NO:64) includes a Ser at amino acidposition 252 instead of a Glu. Similar hybrid molecules have been made.The nucleic acid and amino acid sequences for Met-NLS(signalpeptide)-Gln-rOncFBE6 are set out on SEQ ID NOS:42 and 43. AnotherE6/rOnc molecule is designated Met-Ser-rOncA87FBE6 and is found on SEQID NOS:44 and 45. "A87" refers to the fact that an Ala occurs at aminoacid position 87.

Met-Ser-rOnc-Ang-FBE6 is set out on SEQ ID NOS:46 and 47.

E6FBMet-Ser-rOnc is set out on SEQ ID NOS:48 and 49.

Met-Glu-rOncFBE6 is set out on SEQ ID NOS:50 and 51.

Met-Ser-rOncFBE6 is set out on SEQ ID NOS:50 and 51, with the exceptionthat Ser replaces Glu at amino acid position 2.

MOC31 and MOC162 refer to anti-colon cancer antibodies directed againstthe 17-1-A pancarcinoma antigen which were obtained from Dr. HennieHoogenboom. The Fv region of these antibodies was fused to rOnc. Thenucleic acid and amino acid sequences for MetSerrOnc A87 FBMOC31 are setout on SEQ ID NOS:52 and 53. The nucleic acid and amino acid sequencesfor MOC31FBMetSerrOnc are set out on SEQ ID NOS:54 and 55. The nucleicacid and amino acid sequences for MetSerrOncFBMOC161 are set out on SEQID NOS:56 and 57.

The ligand, IL2 (interleukin 2) was recombinantly fused to rOnc as well.See SEQ ID NOS:58 and 59 for IL2FBMetSerrOnc. See SEQ ID NOS:60 and 61for MetSerrOncFBIL2.

Inhibition of protein synthesis in SF539 cells (which bear thetransferrin receptor) was measured, as described above, for[Met-(-1)Ser]rOnc, E6FB[Met-(-1)Ser]rOnc; [Met-(-1)Ser]rOnc-AngFBE6 and[Met-(-1)Glu]rOncFBE6 constructs and compared with nOnc. The results areshown on Table 3. The three E6 constructs, in particular, had a veryhigh level of activity--up to 45 fold difference over the two non-E6molecules. See also FIG. 5. MetSerOncAng molecule was made correspondingto amino acids 1-107 of SEQ ID NO:47.

                  TABLE 3                                                         ______________________________________                                        Activity of modified rOnc and modified rOncFvs on protein synthesis           RNase             IC.sub.50 (nM)                                                                         Fold Difference                                    ______________________________________                                        nOnc              10        1                                                 [Met-(-1)Ser]rOnc 8        NSD                                                E6FB[Met-(-1)Ser]rOnc                                                                           0.22     45                                                 [Met-(-1)Ser]rOnc-AngFBE6                                                                       0.27     37                                                 [Met-(-1)Glu]rOncFBE6                                                                           0.50     20                                                 ______________________________________                                         The concentrations necessary to inhibit protein synthesis by 50% in SF539     human glioma cells. NSD, no significant difference.                      

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 64                                            - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 104 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..104                                                #/label= nOnc OTHER INFORMATION:                                              #"native ONCONASE (Registered                                                 #from Rana pipiens"emark)                                                     -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 1                                                     #/note= "Xaa = pyroglutamic acid"                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 - Xaa Asp Trp Leu Thr Phe Gln Lys Lys His Il - #e Thr Asn Thr Arg Asp         #                15                                                           - Val Asp Cys Asp Asn Ile Met Ser Thr Asn Le - #u Phe His Cys Lys Asp         #            30                                                               - Lys Asn Thr Phe Ile Tyr Ser Arg Pro Glu Pr - #o Val Lys Ala Ile Cys         #        45                                                                   - Lys Gly Ile Ile Ala Ser Lys Asn Val Leu Th - #r Thr Ser Glu Phe Tyr         #    60                                                                       - Leu Ser Asp Cys Asn Val Thr Ser Arg Pro Cy - #s Lys Tyr Lys Leu Lys         #80                                                                           - Lys Ser Thr Asn Lys Phe Cys Val Thr Cys Gl - #u Asn Gln Ala Pro Val         #                95                                                           - His Phe Val Gly Val Gly Ser Cys                                                         100                                                               - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 83 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..83                                                 #/note= "Rana clone 9 peptide from Rana                                                      pipiens g - #enomic DNA"                                       -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #                15                                                           - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #            30                                                               - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #        45                                                                   - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         #    60                                                                       - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #80                                                                           - Val His Phe                                                                 - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 28 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..28                                                 #/note= "N-terminal sequence of nOnc                                          #in place of pyroglutamic acid in position 1"                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 - Glu Asp Trp Leu Thr Phe Gln Lys Lys His Il - #e Thr Asn Thr Arg Asp         #                15                                                           - Val Asp Cys Asp Asn Ile Met Ser Thr Asn Le - #u Phe                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..34                                                 #/note= "N-terminal sequence of:                                              #eosinophil-derivedmbinant                                                    #(rEDN)"       neurotoxin                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 - Lys Pro Pro Gln Phe Thr Trp Ala Gln Trp Ph - #e Glu Thr Gln His Ile         #                15                                                           - Asn Met Thr Ser Gln Gln Cys Thr Asn Ala Me - #t Gln Val Ile Asn Asn         #            30                                                               - Tyr Gln                                                                     - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 28 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..28                                                 #/note= "N-terminal sequence of:                                                             [Met-(-1)]- #rOncG20, containing a Gly to                                     Asp subst - #itution at position 20 of                                        [Met-(-1)]- #rOnc, and without the extra                       #Met from the E. coliinal                                                     #expression system"erial                                                      -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 - Glu Asp Trp Leu Thr Phe Gln Lys Lys His Il - #e Thr Asn Thr Arg Asp         #                15                                                           - Val Asp Cys Gly Asn Ile Met Ser Thr Asn Le - #u Phe                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..34                                                 #/note= "N-terminal sequence of:                                                             rEDN1-21rOnc - #, without the extra                            #Met from the E. coliinal                                                     #expression system"erial                                                      -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 - Lys Pro Pro Gln Phe Thr Trp Ala Gln Trp Ph - #e Glu Thr Gln His Ile         #                15                                                           - Asn Met Thr Ser Gln Asp Val Asp Cys Asp As - #n Ile Met Ser Thr Asn         #            30                                                               - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Peptide                                                         (B) LOCATION: 1..34                                                 #/note= "N-terminal sequence of:                                                             rEDN1-21rOnc - #G26, containing a Gly to Asp                                  substitution - # at position 26 of                                            rEDN1-21rOnc - #, and without the extra                        #Met from the E. coliinal                                                     #expression system"erial                                                      -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 - Lys Pro Pro Gln Phe Thr Trp Ala Gln Trp Ph - #e Glu Thr Gln His Ile         #                15                                                           - Asn Met Thr Ser Gln Asp Val Asp Cys Gly As - #n Ile Met Ser Thr Asn         #            30                                                               - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 111 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..111                                                #/note= "Frog Lectin from RanaN:                                                             catesbeiana"                                                   -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 - Glu Asn Trp Ala Thr Phe Gln Gln Lys His Il - #e Ile Asn Thr Pro Ile         #                15                                                           - Ile Asn Cys Asn Thr Ile Met Asp Asn Asn Il - #e Tyr Ile Val Gly Gly         #            30                                                               - Gln Cys Lys Arg Val Asn Thr Phe Ile Ile Se - #r Ser Ala Thr Thr Val         #        45                                                                   - Lys Ala Ile Cys Thr Gly Val Ile Asn Met As - #n Val Leu Ser Thr Thr         #    60                                                                       - Arg Phe Gln Leu Asn Thr Cys Thr Arg Thr Se - #r Ile Thr Pro Arg Pro         #80                                                                           - Cys Pro Tyr Ser Ser Arg Thr Glu Thr Asn Ty - #r Ile Cys Val Lys Cys         #                95                                                           - Glu Asn Gln Tyr Pro Val His Phe Ala Gly Il - #e Gly Arg Cys Pro             #           110                                                               - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 134 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..134                                                #/note= "Human eosinophil-derived                                             #(EDN)"        neurotoxin                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 - Lys Pro Pro Gln Phe Thr Trp Ala Gln Trp Ph - #e Glu Thr Gln His Ile         #                15                                                           - Asn Met Thr Ser Gln Gln Cys Thr Asn Ala Me - #t Gln Val Ile Asn Asn         #            30                                                               - Tyr Gln Arg Arg Cys Lys Asn Gln Asn Thr Ph - #e Leu Leu Thr Thr Phe         #        45                                                                   - Ala Asn Val Val Asn Val Cys Gly Asn Pro As - #n Met Thr Cys Pro Ser         #    60                                                                       - Asn Lys Thr Arg Lys Asn Cys His His Ser Gl - #y Ser Gln Val Pro Leu         #80                                                                           - Ile His Cys Asn Leu Thr Thr Pro Ser Pro Gl - #n Asn Ile Ser Asn Cys         #                95                                                           - Arg Tyr Ala Gln Thr Pro Ala Asn Met Phe Ty - #r Ile Val Ala Cys Asp         #           110                                                               - Asn Arg Asp Gln Arg Arg Asp Pro Pro Gln Ty - #r Pro Val Val Pro Val         #       125                                                                   - His Leu Asp Arg Ile Ile                                                         130                                                                       - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 133 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..133                                                #/note= "Human eosinophil cationic                                                           protein ( - #ECP)"                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                - Arg Pro Pro Gln Phe Thr Arg Ala Gln Trp Ph - #e Ala Ile Gln His Ile         #                15                                                           - Ser Leu Asn Pro Pro Arg Cys Thr Ile Ala Me - #t Arg Ala Ile Asn Asn         #            30                                                               - Tyr Arg Trp Arg Cys Lys Asn Gln Asn Thr Ph - #e Leu Arg Thr Thr Phe         #        45                                                                   - Ala Asn Val Val Asn Val Cys Gly Asn Gln Se - #r Ile Arg Cys Pro His         #    60                                                                       - Asn Arg Thr Leu Asn Asn Cys His Arg Ser Ar - #g Phe Arg Val Pro Leu         #80                                                                           - Leu His Cys Asp Leu Ile Asn Pro Gly Ala Gl - #n Asn Ile Ser Asn Cys         #                95                                                           - Arg Tyr Ala Asp Arg Pro Gly Arg Arg Phe Ty - #r Val Val Ala Cys Asp         #           110                                                               - Asn Arg Asp Pro Arg Asp Ser Pro Arg Tyr Pr - #o Val Val Pro Val His         #       125                                                                   - Leu Asp Thr Thr Ile                                                             130                                                                       - (2) INFORMATION FOR SEQ ID NO:11:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 125 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..125                                                #/note= "Bovine angiogenin (Ang)"                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                - Ala Gln Asp Asp Tyr Arg Tyr Ile His Phe Le - #u Thr Gln His Tyr Asp         #                15                                                           - Ala Lys Pro Lys Gly Arg Asn Asp Glu Tyr Cy - #s Phe His Met Met Lys         #            30                                                               - Asn Arg Arg Leu Thr Arg Pro Cys Lys Asp Ar - #g Asn Thr Phe Ile His         #        45                                                                   - Gly Asn Lys Asn Asp Ile Lys Ala Ile Cys Gl - #u Asp Arg Asn Gly Gln         #    60                                                                       - Pro Tyr Arg Gly Asp Leu Arg Ile Ser Lys Se - #r Glu Phe Gln Ile Thr         #80                                                                           - Ile Cys Lys His Lys Gly Gly Ser Ser Arg Pr - #o Pro Cys Arg Tyr Gly         #                95                                                           - Ala Thr Glu Asp Ser Arg Val Ile Val Val Gl - #y Cys Glu Asn Gly Leu         #           110                                                               - Pro Val His Phe Asp Glu Ser Phe Ile Thr Pr - #o Arg His                     #       125                                                                   - (2) INFORMATION FOR SEQ ID NO:12:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 124 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..124                                                #/note= "Bovine seminal RNase"N:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                - Lys Glu Ser Ala Ala Ala Lys Phe Glu Arg Gl - #n His Met Asp Ser Gly         #                15                                                           - Asn Ser Pro Ser Ser Ser Ser Asn Tyr Cys As - #n Leu Met Met Cys Cys         #            30                                                               - Arg Lys Met Thr Gln Gly Lys Cys Lys Pro Va - #l Asn Thr Phe Val His         #        45                                                                   - Glu Ser Leu Ala Asp Val Lys Ala Val Cys Se - #r Gln Lys Lys Val Thr         #    60                                                                       - Cys Lys Asn Gly Gln Thr Asn Cys Tyr Gln Se - #r Lys Ser Thr Met Arg         #80                                                                           - Ile Thr Asp Cys Arg Glu Thr Gly Ser Ser Ly - #s Tyr Pro Asn Cys Ala         #                95                                                           - Tyr Lys Thr Thr Gln Val Glu Lys His Ile Il - #e Val Ala Cys Gly Gly         #           110                                                               - Lys Pro Ser Val Pro Val His Phe Asp Ala Se - #r Val                         #       120                                                                   - (2) INFORMATION FOR SEQ ID NO:13:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 124 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..124                                                #/note= "Bovine pancreatic RNase A"                                           -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                - Lys Glu Thr Ala Ala Ala Lys Phe Glu Arg Gl - #n His Met Asp Ser Ser         #                15                                                           - Thr Ser Ala Ala Ser Ser Ser Asn Tyr Cys As - #n Gln Met Met Lys Ser         #            30                                                               - Arg Asn Leu Thr Lys Asp Arg Cys Lys Pro Va - #l Asn Thr Phe Val His         #        45                                                                   - Glu Ser Leu Ala Asp Val Gln Ala Val Cys Se - #r Gln Lys Asn Val Ala         #    60                                                                       - Cys Lys Asn Gly Gln Thr Asn Cys Tyr Gln Se - #r Tyr Ser Thr Met Ser         #80                                                                           - Ile Thr Asp Cys Arg Glu Thr Gly Ser Ser Ly - #s Tyr Pro Asn Cys Ala         #                95                                                           - Tyr Lys Thr Thr Gln Ala Asn Lys His Ile Il - #e Val Ala Cys Glu Gly         #           110                                                               - Asn Pro Val Val Pro Val His Phe Asp Ala Se - #r Val                         #       120                                                                   - (2) INFORMATION FOR SEQ ID NO:14:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 3                                                     #/note= "Xaa = Ser, Tyr or Thr":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                - Met Lys Xaa Pro                                                             - (2) INFORMATION FOR SEQ ID NO:15:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 4                                                     #/note= "Xaa = Ser, Tyr or Thr":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                - Met Lys Pro Xaa                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:16:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 3                                                     #/note= "Xaa = Ser, Tyr or Thr":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                - Met Asn Xaa Pro                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:17:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 2                                                     #/note= "Xaa = Ser, Tyr or Thr":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                - Met Xaa Lys Pro                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:18:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 2                                                     #/note= "Xaa = Ser, Tyr or Thr":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                - Met Xaa Pro Lys                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:19:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 321 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..321                                                #/note= "MetSerOnc99Ang117"TION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                - ATC TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Ile Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          #        321T GTT CAG TCA ATT TTC CGT CGT CC - #G                             Val His Phe Val Gln Ser Ile Phe Arg Arg Pr - #o                               #           105                                                               - (2) INFORMATION FOR SEQ ID NO:20:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 107 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                - Ile Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gln Ser Ile Phe Arg Arg Pr - #o                             #           105                                                               - (2) INFORMATION FOR SEQ ID NO:21:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 333 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..333                                                #/note= "EDNGlyOnc" INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                - ATG AAA CCG CCG CAG TTC ACT TGG GCT CAG TG - #G TTC GAA ACT CAG CAT           48                                                                          Met Lys Pro Pro Gln Phe Thr Trp Ala Gln Tr - #p Phe Glu Thr Gln His           #                 15                                                          - ATC AAC ATG ACT TCT CAG GAT GTT GAT TGT GG - #T AAT ATC ATG TCA ACA           96                                                                          Ile Asn Met Thr Ser Gln Asp Val Asp Cys Gl - #y Asn Ile Met Ser Thr           #             30                                                              - AAC TTG TTC CAC TGC AAG GAC AAG AAC ACT TT - #T ATC TAT TCA CGT CCT          144                                                                          Asn Leu Phe His Cys Lys Asp Lys Asn Thr Ph - #e Ile Tyr Ser Arg Pro           #         45                                                                  - GAG CCA GTG AAG GCC ATC TGT AAA GGA ATT AT - #A GCC TCC AAA AAT GTG          192                                                                          Glu Pro Val Lys Ala Ile Cys Lys Gly Ile Il - #e Ala Ser Lys Asn Val           #     60                                                                      - TTA ACT ACC TCT GAG TTT TAT CTC TCT GAT TG - #C AAT GTA ACA AGC AGG          240                                                                          Leu Thr Thr Ser Glu Phe Tyr Leu Ser Asp Cy - #s Asn Val Thr Ser Arg           # 80                                                                          - CCT TGC AAG TAT AAA TTA AAG AAA TCA ACT AA - #T AAA TTT TGT GTA ACT          288                                                                          Pro Cys Lys Tyr Lys Leu Lys Lys Ser Thr As - #n Lys Phe Cys Val Thr           #                 95                                                          - TGT GAA AAT CAG GCA CCA GTT CAT TTT GTT GG - #A GTT GGA TCT TGT              33 - #3                                                                      Cys Glu Asn Gln Ala Pro Val His Phe Val Gl - #y Val Gly Ser Cys               #           110                                                               - (2) INFORMATION FOR SEQ ID NO:22:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 111 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                - Met Lys Pro Pro Gln Phe Thr Trp Ala Gln Tr - #p Phe Glu Thr Gln His         #                 15                                                          - Ile Asn Met Thr Ser Gln Asp Val Asp Cys Gl - #y Asn Ile Met Ser Thr         #             30                                                              - Asn Leu Phe His Cys Lys Asp Lys Asn Thr Ph - #e Ile Tyr Ser Arg Pro         #         45                                                                  - Glu Pro Val Lys Ala Ile Cys Lys Gly Ile Il - #e Ala Ser Lys Asn Val         #     60                                                                      - Leu Thr Thr Ser Glu Phe Tyr Leu Ser Asp Cy - #s Asn Val Thr Ser Arg         # 80                                                                          - Pro Cys Lys Tyr Lys Leu Lys Lys Ser Thr As - #n Lys Phe Cys Val Thr         #                 95                                                          - Cys Glu Asn Gln Ala Pro Val His Phe Val Gl - #y Val Gly Ser Cys             #           110                                                               - (2) INFORMATION FOR SEQ ID NO:23:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 315 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..315                                                #/note= "MetTyrrOnc"INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                - ATG TAT GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Tyr Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ACT TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Thr Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          #            315   GA GTT GGA TCT TGT                                         Val His Phe Val Gly Val Gly Ser Cys                                           #           105                                                               - (2) INFORMATION FOR SEQ ID NO:24:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 105 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                - Met Tyr Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Thr Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys                                         #           105                                                               - (2) INFORMATION FOR SEQ ID NO:25:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 315 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..315                                                #/note= "MetSerrOnc"INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ACT TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Thr Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          #            315   GA GTT GGA TCT TGT                                         Val His Phe Val Gly Val Gly Ser Cys                                           #           105                                                               - (2) INFORMATION FOR SEQ ID NO:26:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 105 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Thr Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys                                         #           105                                                               - (2) INFORMATION FOR SEQ ID NO:27:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 318 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..318                                                #/note= "MetLysTyrrOnc"ORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                - ATG AAA TAT GAT TGG CTT ACA TTT CAG AAA AA - #A CAC ATC ACA AAC ACA           48                                                                          Met Lys Tyr Asp Trp Leu Thr Phe Gln Lys Ly - #s His Ile Thr Asn Thr           #                 15                                                          - AGG GAT GTT GAT TGT GAT AAT ATC ATG TCA AC - #A AAC TTG TTC CAC TGC           96                                                                          Arg Asp Val Asp Cys Asp Asn Ile Met Ser Th - #r Asn Leu Phe His Cys           #             30                                                              - AAG GAC AAG AAC ACT TTT ATC TAT TCA CGT CC - #T GAG CCA GTG AAG GCC          144                                                                          Lys Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pr - #o Glu Pro Val Lys Ala           #         45                                                                  - ATC TGT AAA GGA ATT ATA GCC TCC AAA AAT GT - #G TTA ACT ACC TCT GAG          192                                                                          Ile Cys Lys Gly Ile Ile Ala Ser Lys Asn Va - #l Leu Thr Thr Ser Glu           #     60                                                                      - TTT TAT CTC TCT GAT TGC AAT GTA ACA AGC AG - #G CCT TGC AAG TAT AAA          240                                                                          Phe Tyr Leu Ser Asp Cys Asn Val Thr Ser Ar - #g Pro Cys Lys Tyr Lys           # 80                                                                          - TTA AAG AAA TCA ACT AAT AAA TTT TGT GTA AC - #T TGT GAA AAT CAG GCA          288                                                                          Leu Lys Lys Ser Thr Asn Lys Phe Cys Val Th - #r Cys Glu Asn Gln Ala           #                 95                                                          #          318     TT GGA GTT GGA TCT TGT                                     Pro Val His Phe Val Gly Val Gly Ser Cys                                       #           105                                                               - (2) INFORMATION FOR SEQ ID NO:28:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 106 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                - Met Lys Tyr Asp Trp Leu Thr Phe Gln Lys Ly - #s His Ile Thr Asn Thr         #                 15                                                          - Arg Asp Val Asp Cys Asp Asn Ile Met Ser Th - #r Asn Leu Phe His Cys         #             30                                                              - Lys Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pr - #o Glu Pro Val Lys Ala         #         45                                                                  - Ile Cys Lys Gly Ile Ile Ala Ser Lys Asn Va - #l Leu Thr Thr Ser Glu         #     60                                                                      - Phe Tyr Leu Ser Asp Cys Asn Val Thr Ser Ar - #g Pro Cys Lys Tyr Lys         # 80                                                                          - Leu Lys Lys Ser Thr Asn Lys Phe Cys Val Th - #r Cys Glu Asn Gln Ala         #                 95                                                          - Pro Val His Phe Val Gly Val Gly Ser Cys                                     #           105                                                               - (2) INFORMATION FOR SEQ ID NO:29:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 321 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..321                                                #/note= "MetAlaAlaTyrrOnc"ATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                - ATG GCT GCT TAT GAT TGG CTT ACA TTT CAG AA - #A AAA CAC ATC ACA AAC           48                                                                          Met Ala Ala Tyr Asp Trp Leu Thr Phe Gln Ly - #s Lys His Ile Thr Asn           #                 15                                                          - ACA AGG GAT GTT GAT TGT GAT AAT ATC ATG TC - #A ACA AAC TTG TTC CAC           96                                                                          Thr Arg Asp Val Asp Cys Asp Asn Ile Met Se - #r Thr Asn Leu Phe His           #             30                                                              - TGC AAG GAC AAG AAC ACT TTT ATC TAT TCA CG - #T CCT GAG CCA GTG AAG          144                                                                          Cys Lys Asp Lys Asn Thr Phe Ile Tyr Ser Ar - #g Pro Glu Pro Val Lys           #         45                                                                  - GCC ATC TGT AAA GGA ATT ATA GCC TCC AAA AA - #T GTG TTA ACT ACC TCT          192                                                                          Ala Ile Cys Lys Gly Ile Ile Ala Ser Lys As - #n Val Leu Thr Thr Ser           #     60                                                                      - GAG TTT TAT CTC TCT GAT TGC AAT GTA ACA AG - #C AGG CCT TGC AAG TAT          240                                                                          Glu Phe Tyr Leu Ser Asp Cys Asn Val Thr Se - #r Arg Pro Cys Lys Tyr           # 80                                                                          - AAA TTA AAG AAA TCA ACT AAT AAA TTT TGT GT - #A ACT TGT GAA AAT CAG          288                                                                          Lys Leu Lys Lys Ser Thr Asn Lys Phe Cys Va - #l Thr Cys Glu Asn Gln           #                 95                                                          #        321T CAT TTT GTT GGA GTT GGA TCT TG - #T                             Ala Pro Val His Phe Val Gly Val Gly Ser Cy - #s                               #           105                                                               - (2) INFORMATION FOR SEQ ID NO:30:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 107 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                - Met Ala Ala Tyr Asp Trp Leu Thr Phe Gln Ly - #s Lys His Ile Thr Asn         #                 15                                                          - Thr Arg Asp Val Asp Cys Asp Asn Ile Met Se - #r Thr Asn Leu Phe His         #             30                                                              - Cys Lys Asp Lys Asn Thr Phe Ile Tyr Ser Ar - #g Pro Glu Pro Val Lys         #         45                                                                  - Ala Ile Cys Lys Gly Ile Ile Ala Ser Lys As - #n Val Leu Thr Thr Ser         #     60                                                                      - Glu Phe Tyr Leu Ser Asp Cys Asn Val Thr Se - #r Arg Pro Cys Lys Tyr         # 80                                                                          - Lys Leu Lys Lys Ser Thr Asn Lys Phe Cys Va - #l Thr Cys Glu Asn Gln         #                 95                                                          - Ala Pro Val His Phe Val Gly Val Gly Ser Cy - #s                             #           105                                                               - (2) INFORMATION FOR SEQ ID NO:31:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 336 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..336                                                #/note= "NLSMetSerrOnc"ORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                - CCC AAG AAG AAG CGG AAG GTG ATG TCA GAT TG - #G CTT ACA TTT CAG AAA           48                                                                          Pro Lys Lys Lys Arg Lys Val Met Ser Asp Tr - #p Leu Thr Phe Gln Lys           #                 15                                                          - AAA CAC ATC ACA AAC ACA AGG GAT GTT GAT TG - #T GAT AAT ATC ATG TCA           96                                                                          Lys His Ile Thr Asn Thr Arg Asp Val Asp Cy - #s Asp Asn Ile Met Ser           #             30                                                              - ACA AAC TTG TTC CAC TGC AAG GAC AAG AAC AC - #T TTT ATC TAT TCA CGT          144                                                                          Thr Asn Leu Phe His Cys Lys Asp Lys Asn Th - #r Phe Ile Tyr Ser Arg           #         45                                                                  - CCT GAG CCA GTG AAG GCC ATC TGT AAA GGA AT - #T ATA GCC TCC AAA AAT          192                                                                          Pro Glu Pro Val Lys Ala Ile Cys Lys Gly Il - #e Ile Ala Ser Lys Asn           #     60                                                                      - GTG TTA ACT ACC TCT GAG TTT TAT CTC TCT GA - #T TGC AAT GTA ACA AGC          240                                                                          Val Leu Thr Thr Ser Glu Phe Tyr Leu Ser As - #p Cys Asn Val Thr Ser           # 80                                                                          - AGG CCT TGC AAG TAT AAA TTA AAG AAA TCA AC - #T AAT AAA TTT TGT GTA          288                                                                          Arg Pro Cys Lys Tyr Lys Leu Lys Lys Ser Th - #r Asn Lys Phe Cys Val           #                 95                                                          - ACT TGT GAA AAT CAG GCA CCA GTT CAT TTT GT - #T GGA GTT GGA TCT TGT          336                                                                          Thr Cys Glu Asn Gln Ala Pro Val His Phe Va - #l Gly Val Gly Ser Cys           #           110                                                               - (2) INFORMATION FOR SEQ ID NO:32:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 112 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                - Pro Lys Lys Lys Arg Lys Val Met Ser Asp Tr - #p Leu Thr Phe Gln Lys         #                 15                                                          - Lys His Ile Thr Asn Thr Arg Asp Val Asp Cy - #s Asp Asn Ile Met Ser         #             30                                                              - Thr Asn Leu Phe His Cys Lys Asp Lys Asn Th - #r Phe Ile Tyr Ser Arg         #         45                                                                  - Pro Glu Pro Val Lys Ala Ile Cys Lys Gly Il - #e Ile Ala Ser Lys Asn         #     60                                                                      - Val Leu Thr Thr Ser Glu Phe Tyr Leu Ser As - #p Cys Asn Val Thr Ser         # 80                                                                          - Arg Pro Cys Lys Tyr Lys Leu Lys Lys Ser Th - #r Asn Lys Phe Cys Val         #                 95                                                          - Thr Cys Glu Asn Gln Ala Pro Val His Phe Va - #l Gly Val Gly Ser Cys         #           110                                                               - (2) INFORMATION FOR SEQ ID NO:33:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 2                                                     #/note= "Xaa = an aliphatic amino acid,                                                      Ala, Leu, - # Ile, Val, or Pro"                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 3                                                     #/note= "Xaa = an aliphatic amino acid,                                                      Ala, Leu, - # Ile, Val or Pro"                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: Modified-sit - #e                                               (B) LOCATION: 4                                                     #/note= "Xaa = Ser, Met, Cys, Ala or                                                         Gln"                                                           -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                - Cys Xaa Xaa Xaa                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:34:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 4 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                - Cys Val Ile Met                                                             1                                                                             - (2) INFORMATION FOR SEQ ID NO:35:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                #             27   ATAA YATCATG                                               - (2) INFORMATION FOR SEQ ID NO:36:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 27 base                                                           (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                #             27   CWGT KCAYTTT                                               - (2) INFORMATION FOR SEQ ID NO:37:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 249 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY:                                                                 (B) LOCATION: 1..249                                                #/note= "Rana 9"HER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                - GATGTTGATT GTGATAATAT CATGTCAACA AACTTGTTCC ACTGCAAGGA CA - #AGAACACT         60                                                                          - TTTATCTATT CACGTCCTGA GCCAGTGAAG GCCATCTGTA AAGGAATTAT AG - #CCTCCAAA        120                                                                          - AATGTGTTAA CTACCTCTGA GTTTTATCTC TCTGATTGCA ATGTAACAAG CA - #GGCCTTGC        180                                                                          - AAGTATAAAT TAAAGAAATC AACTAATAAA TTTTGTGTAA CTTGTGAAAA TC - #AGGCACCA        240                                                                          #        249                                                                  - (2) INFORMATION FOR SEQ ID NO:38:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 315 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..315                                                #/note= "[Met-(-1)]rOnc"RMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                - ATG GAG GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Glu Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          #            315   GA GTT GGA TCT TGT                                         Val His Phe Val Gly Val Gly Ser Cys                                           #           105                                                               - (2) INFORMATION FOR SEQ ID NO:39:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 105 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                - Met Glu Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys                                         #           105                                                               - (2) INFORMATION FOR SEQ ID NO:40:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1065 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1065                                               #/note= "sFvFBMetGluOnc"RMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                - GAC ATC AAG ATG ACC CAG TCT CCA TCT TCC AT - #G TAT GCA TCT CTA GGA           48                                                                          Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Me - #t Tyr Ala Ser Leu Gly           #                 15                                                          - GAG AGA GTC ACT TTC ACT TGC AAG GCG AGT CA - #G GAC ATT AAT AAC TAT           96                                                                          Glu Arg Val Thr Phe Thr Cys Lys Ala Ser Gl - #n Asp Ile Asn Asn Tyr           #             30                                                              - TTA TGC TGG TTC CAG CAG AAA CCA GGG AAA TC - #T CCT AAG ACC CTG ATC          144                                                                          Leu Cys Trp Phe Gln Gln Lys Pro Gly Lys Se - #r Pro Lys Thr Leu Ile           #         45                                                                  - TAT CGT GCA AAC AGA CTG GTA GAT GGG GTC CC - #A TCA AGG TTC AGT GGC          192                                                                          Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pr - #o Ser Arg Phe Ser Gly           #     60                                                                      - AGT GGA TCT GGA CAA GAT TAT TCT CTC ACC AT - #T AGC AGC CTG GAG TAT          240                                                                          Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Il - #e Ser Ser Leu Glu Tyr           # 80                                                                          - GAA GAT ATG GGA ATT TAT TAT TGT CTA CAG TA - #T GAT GAG TTT CCG TAC          288                                                                          Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Ty - #r Asp Glu Phe Pro Tyr           #                 95                                                          - ACG TTC GGA GGG GGG ACC AAG CTG GAA ATA AA - #A GGA GGC GGT GGC TCG          336                                                                          Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Ly - #s Gly Gly Gly Gly Ser           #           110                                                               - GGC GGT GGC GGA TCG GGT GGC GGC GGC TCT GA - #G GTT CAG CTC CAG CAG          384                                                                          Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #u Val Gln Leu Gln Gln           #       125                                                                   - TCT GGG ACT GTA CTG GCA AGG CCT GGG GCT TC - #A GTG AAG ATG TCC TGC          432                                                                          Ser Gly Thr Val Leu Ala Arg Pro Gly Ala Se - #r Val Lys Met Ser Cys           #   140                                                                       - AAG GCT TCT GGC TAC ACC TTT TCC AGC TAC TG - #G ATG CAC TGG ATA AAA          480                                                                          Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Tr - #p Met His Trp Ile Lys           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - CAG AGG CCT GGA CAG GGT CTG GAC TGG ATT GT - #C GCT ATT GAT CCT CGA          528                                                                          Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile Va - #l Ala Ile Asp Pro Arg           #               175                                                           - AAT AGT GAT ACT ATT TAC AAC CCG CAA TTC AA - #A CAC AAG GCC AAA CTG          576                                                                          Asn Ser Asp Thr Ile Tyr Asn Pro Gln Phe Ly - #s His Lys Ala Lys Leu           #           190                                                               - ACT GCA GTC ACC TCC ACC AGC ACT GCC TAC AT - #G GAA CTC AAC AGC CTG          624                                                                          Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr Me - #t Glu Leu Asn Ser Leu           #       205                                                                   - ACA AAT GAG GAC TCT GCG GTC TAT TAC TGT AC - #C CCT CTT TAT TAC TTT          672                                                                          Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys Th - #r Pro Leu Tyr Tyr Phe           #   220                                                                       - GAC TCC TGG GGC CAA GGC ACC ACT CTC ACA GT - #C TCC TCA GCC AAG AAA          720                                                                          Asp Ser Trp Gly Gln Gly Thr Thr Leu Thr Va - #l Ser Ser Ala Lys Lys           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - CTG AAC GAC GCT CAG GCG CCG AAG AGT GAT AT - #G GAG GAT TGG CTT ACA          768                                                                          Leu Asn Asp Ala Gln Ala Pro Lys Ser Asp Me - #t Glu Asp Trp Leu Thr           #               255                                                           - TTT CAG AAA AAA CAC ATC ACA AAC ACA AGG GA - #T GTT GAT TGT GAT AAT          816                                                                          Phe Gln Lys Lys His Ile Thr Asn Thr Arg As - #p Val Asp Cys Asp Asn           #           270                                                               - ATC ATG TCA ACA AAC TTG TTC CAC TGC AAG GA - #C AAG AAC ACT TTT ATC          864                                                                          Ile Met Ser Thr Asn Leu Phe His Cys Lys As - #p Lys Asn Thr Phe Ile           #       285                                                                   - TAT TCA CGT CCT GAG CCA GTG AAG GCC ATC TG - #T AAA GGA ATT ATA GCC          912                                                                          Tyr Ser Arg Pro Glu Pro Val Lys Ala Ile Cy - #s Lys Gly Ile Ile Ala           #   300                                                                       - TCC AAA AAT GTG TTA ACT ACC TCT GAG TTT TA - #T CTC TCT GAT TGC AAT          960                                                                          Ser Lys Asn Val Leu Thr Thr Ser Glu Phe Ty - #r Leu Ser Asp Cys Asn           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - GTA ACA AGC AGG CCT TGC AAG TAT AAA TTA AA - #G AAA TCA ACT AAT AAA         1008                                                                          Val Thr Ser Arg Pro Cys Lys Tyr Lys Leu Ly - #s Lys Ser Thr Asn Lys           #               335                                                           - TTT TGT GTA ACT TGT GAA AAT CAG GCA CCA GT - #T CAT TTT GTT GGA GTT         1056                                                                          Phe Cys Val Thr Cys Glu Asn Gln Ala Pro Va - #l His Phe Val Gly Val           #           350                                                               #       1065                                                                  Gly Ser Cys                                                                           355                                                                   - (2) INFORMATION FOR SEQ ID NO:41:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 355 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                - Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Me - #t Tyr Ala Ser Leu Gly         #                 15                                                          - Glu Arg Val Thr Phe Thr Cys Lys Ala Ser Gl - #n Asp Ile Asn Asn Tyr         #             30                                                              - Leu Cys Trp Phe Gln Gln Lys Pro Gly Lys Se - #r Pro Lys Thr Leu Ile         #         45                                                                  - Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pr - #o Ser Arg Phe Ser Gly         #     60                                                                      - Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Il - #e Ser Ser Leu Glu Tyr         # 80                                                                          - Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Ty - #r Asp Glu Phe Pro Tyr         #                 95                                                          - Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Ly - #s Gly Gly Gly Gly Ser         #           110                                                               - Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #u Val Gln Leu Gln Gln         #       125                                                                   - Ser Gly Thr Val Leu Ala Arg Pro Gly Ala Se - #r Val Lys Met Ser Cys         #   140                                                                       - Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Tr - #p Met His Trp Ile Lys         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile Va - #l Ala Ile Asp Pro Arg         #               175                                                           - Asn Ser Asp Thr Ile Tyr Asn Pro Gln Phe Ly - #s His Lys Ala Lys Leu         #           190                                                               - Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr Me - #t Glu Leu Asn Ser Leu         #       205                                                                   - Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys Th - #r Pro Leu Tyr Tyr Phe         #   220                                                                       - Asp Ser Trp Gly Gln Gly Thr Thr Leu Thr Va - #l Ser Ser Ala Lys Lys         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Leu Asn Asp Ala Gln Ala Pro Lys Ser Asp Me - #t Glu Asp Trp Leu Thr         #               255                                                           - Phe Gln Lys Lys His Ile Thr Asn Thr Arg As - #p Val Asp Cys Asp Asn         #           270                                                               - Ile Met Ser Thr Asn Leu Phe His Cys Lys As - #p Lys Asn Thr Phe Ile         #       285                                                                   - Tyr Ser Arg Pro Glu Pro Val Lys Ala Ile Cy - #s Lys Gly Ile Ile Ala         #   300                                                                       - Ser Lys Asn Val Leu Thr Thr Ser Glu Phe Ty - #r Leu Ser Asp Cys Asn         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Val Thr Ser Arg Pro Cys Lys Tyr Lys Leu Ly - #s Lys Ser Thr Asn Lys         #               335                                                           - Phe Cys Val Thr Cys Glu Asn Gln Ala Pro Va - #l His Phe Val Gly Val         #           350                                                               - Gly Ser Cys                                                                         355                                                                   - (2) INFORMATION FOR SEQ ID NO:42:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1137 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1137                                               #/note= "SigPepGlnOncFBE6"ATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                - ATG GGT CTG GAA AAA TCT CTG ATC CTG TTC CC - #G CTG TTC TTC CTG CTG           48                                                                          Met Gly Leu Glu Lys Ser Leu Ile Leu Phe Pr - #o Leu Phe Phe Leu Leu           #                 15                                                          - CTG GGT TGG GTT CAG CCG TCT CTG GGT CAG GA - #T TGG CTT ACA TTT CAG           96                                                                          Leu Gly Trp Val Gln Pro Ser Leu Gly Gln As - #p Trp Leu Thr Phe Gln           #             30                                                              - AAA AAA CAC ATC ACA AAC ACA AGG GAT GTT GA - #T TGT GAT AAT ATC ATG          144                                                                          Lys Lys His Ile Thr Asn Thr Arg Asp Val As - #p Cys Asp Asn Ile Met           #         45                                                                  - TCA ACA AAC TTG TTC CAC TGC AAG GAC AAG AA - #C ACT TTT ATC TAT TCA          192                                                                          Ser Thr Asn Leu Phe His Cys Lys Asp Lys As - #n Thr Phe Ile Tyr Ser           #     60                                                                      - CGT CCT GAG CCA GTG AAG GCC ATC TGT AAA GG - #A ATT ATA GCC TCC AAA          240                                                                          Arg Pro Glu Pro Val Lys Ala Ile Cys Lys Gl - #y Ile Ile Ala Ser Lys           # 80                                                                          - AAT GTG TTA ACT ACC TCT GAG TTT TAT CTC TC - #T GAT TGC AAT GTA ACA          288                                                                          Asn Val Leu Thr Thr Ser Glu Phe Tyr Leu Se - #r Asp Cys Asn Val Thr           #                 95                                                          - AGC AGG CCT TGC AAG TAT AAA TTA AAG AAA TC - #A ACT AAT AAA TTT TGT          336                                                                          Ser Arg Pro Cys Lys Tyr Lys Leu Lys Lys Se - #r Thr Asn Lys Phe Cys           #           110                                                               - GTA ACT TGT GAA AAT CAG GCA CCA GTT CAT TT - #T GTT GGA GTT GGA TCT          384                                                                          Val Thr Cys Glu Asn Gln Ala Pro Val His Ph - #e Val Gly Val Gly Ser           #       125                                                                   - TGT GCC AAG AAA CTG AAC GAC GCT CAG GCG CC - #G AAG AGT GAT GAC ATC          432                                                                          Cys Ala Lys Lys Leu Asn Asp Ala Gln Ala Pr - #o Lys Ser Asp Asp Ile           #   140                                                                       - AAG ATG ACC CAG TCT CCA TCT TCC ATG TAT GC - #A TCT CTA GGA GAG AGA          480                                                                          Lys Met Thr Gln Ser Pro Ser Ser Met Tyr Al - #a Ser Leu Gly Glu Arg           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - GTC ACT TTC ACT TGC AAG GCG AGT CAG GAC AT - #T AAT AAC TAT TTA TGC          528                                                                          Val Thr Phe Thr Cys Lys Ala Ser Gln Asp Il - #e Asn Asn Tyr Leu Cys           #               175                                                           - TGG TTC CAG CAG AAA CCA GGG AAA TCT CCT AA - #G ACC CTG ATC TAT CGT          576                                                                          Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Ly - #s Thr Leu Ile Tyr Arg           #           190                                                               - GCA AAC AGA CTG GTA GAT GGG GTC CCA TCA AG - #G TTC AGT GGC AGT GGA          624                                                                          Ala Asn Arg Leu Val Asp Gly Val Pro Ser Ar - #g Phe Ser Gly Ser Gly           #       205                                                                   - TCT GGA CAA GAT TAT TCT CTC ACC ATT AGC AG - #C CTG GAG TAT GAA GAT          672                                                                          Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Se - #r Leu Glu Tyr Glu Asp           #   220                                                                       - ATG GGA ATT TAT TAT TGT CTA CAG TAT GAT GA - #G TTT CCG TAC ACG TTC          720                                                                          Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Gl - #u Phe Pro Tyr Thr Phe           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GGA GGG GGG ACC AAG CTG GAA ATA AAA GGA GG - #C GGT GGC TCG GGC GGT          768                                                                          Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gl - #y Gly Gly Ser Gly Gly           #               255                                                           - GGC GGA TCG GGT GGC GGC GGC TCT GAG GTT CA - #G CTC CAG CAG TCT GGG          816                                                                          Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gl - #n Leu Gln Gln Ser Gly           #           270                                                               - ACT GTA CTG GCA AGG CCT GGG GCT TCA GTG AA - #G ATG TCC TGC AAG GCT          864                                                                          Thr Val Leu Ala Arg Pro Gly Ala Ser Val Ly - #s Met Ser Cys Lys Ala           #       285                                                                   - TCT GGC TAC ACC TTT TCC AGC TAC TGG ATG CA - #C TGG ATA AAA CAG AGG          912                                                                          Ser Gly Tyr Thr Phe Ser Ser Tyr Trp Met Hi - #s Trp Ile Lys Gln Arg           #   300                                                                       - CCT GGA CAG GGT CTG GAC TGG ATT GTC GCT AT - #T GAT CCT CGA AAT AGT          960                                                                          Pro Gly Gln Gly Leu Asp Trp Ile Val Ala Il - #e Asp Pro Arg Asn Ser           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - GAT ACT ATT TAC AAC CCG CAA TTC AAA CAC AA - #G GCC AAA CTG ACT GCA         1008                                                                          Asp Thr Ile Tyr Asn Pro Gln Phe Lys His Ly - #s Ala Lys Leu Thr Ala           #               335                                                           - GTC ACC TCC ACC AGC ACT GCC TAC ATG GAA CT - #C AAC AGC CTG ACA AAT         1056                                                                          Val Thr Ser Thr Ser Thr Ala Tyr Met Glu Le - #u Asn Ser Leu Thr Asn           #           350                                                               - GAG GAC TCT GCG GTC TAT TAC TGT ACC CCT CT - #T TAT TAC TTT GAC TCC         1104                                                                          Glu Asp Ser Ala Val Tyr Tyr Cys Thr Pro Le - #u Tyr Tyr Phe Asp Ser           #       365                                                                   #       1137A GGC ACC ACT CTC ACA GTC TCC TC - #A                             Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Se - #r                               #   375                                                                       - (2) INFORMATION FOR SEQ ID NO:43:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 379 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                - Met Gly Leu Glu Lys Ser Leu Ile Leu Phe Pr - #o Leu Phe Phe Leu Leu         #                 15                                                          - Leu Gly Trp Val Gln Pro Ser Leu Gly Gln As - #p Trp Leu Thr Phe Gln         #             30                                                              - Lys Lys His Ile Thr Asn Thr Arg Asp Val As - #p Cys Asp Asn Ile Met         #         45                                                                  - Ser Thr Asn Leu Phe His Cys Lys Asp Lys As - #n Thr Phe Ile Tyr Ser         #     60                                                                      - Arg Pro Glu Pro Val Lys Ala Ile Cys Lys Gl - #y Ile Ile Ala Ser Lys         # 80                                                                          - Asn Val Leu Thr Thr Ser Glu Phe Tyr Leu Se - #r Asp Cys Asn Val Thr         #                 95                                                          - Ser Arg Pro Cys Lys Tyr Lys Leu Lys Lys Se - #r Thr Asn Lys Phe Cys         #           110                                                               - Val Thr Cys Glu Asn Gln Ala Pro Val His Ph - #e Val Gly Val Gly Ser         #       125                                                                   - Cys Ala Lys Lys Leu Asn Asp Ala Gln Ala Pr - #o Lys Ser Asp Asp Ile         #   140                                                                       - Lys Met Thr Gln Ser Pro Ser Ser Met Tyr Al - #a Ser Leu Gly Glu Arg         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val Thr Phe Thr Cys Lys Ala Ser Gln Asp Il - #e Asn Asn Tyr Leu Cys         #               175                                                           - Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Ly - #s Thr Leu Ile Tyr Arg         #           190                                                               - Ala Asn Arg Leu Val Asp Gly Val Pro Ser Ar - #g Phe Ser Gly Ser Gly         #       205                                                                   - Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Se - #r Leu Glu Tyr Glu Asp         #   220                                                                       - Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Gl - #u Phe Pro Tyr Thr Phe         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gl - #y Gly Gly Ser Gly Gly         #               255                                                           - Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gl - #n Leu Gln Gln Ser Gly         #           270                                                               - Thr Val Leu Ala Arg Pro Gly Ala Ser Val Ly - #s Met Ser Cys Lys Ala         #       285                                                                   - Ser Gly Tyr Thr Phe Ser Ser Tyr Trp Met Hi - #s Trp Ile Lys Gln Arg         #   300                                                                       - Pro Gly Gln Gly Leu Asp Trp Ile Val Ala Il - #e Asp Pro Arg Asn Ser         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Asp Thr Ile Tyr Asn Pro Gln Phe Lys His Ly - #s Ala Lys Leu Thr Ala         #               335                                                           - Val Thr Ser Thr Ser Thr Ala Tyr Met Glu Le - #u Asn Ser Leu Thr Asn         #           350                                                               - Glu Asp Ser Ala Val Tyr Tyr Cys Thr Pro Le - #u Tyr Tyr Phe Asp Ser         #       365                                                                   - Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Se - #r                             #   375                                                                       - (2) INFORMATION FOR SEQ ID NO:44:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1074 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1074                                               #/note= "MetSerOncA87FBE6"ATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT GCT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Ala Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          - GTT CAT TTT GTT GGA GTT GGA TCT TGT GCC AA - #G AAA CTG AAC GAC GCT          336                                                                          Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala           #           110                                                               - CAG GCG CCG AAG AGT GAT GAC ATC AAG ATG AC - #C CAG TCT CCA TCT TCC          384                                                                          Gln Ala Pro Lys Ser Asp Asp Ile Lys Met Th - #r Gln Ser Pro Ser Ser           #       125                                                                   - ATG TAT GCA TCT CTA GGA GAG AGA GTC ACT TT - #C ACT TGC AAG GCG AGT          432                                                                          Met Tyr Ala Ser Leu Gly Glu Arg Val Thr Ph - #e Thr Cys Lys Ala Ser           #   140                                                                       - CAG GAC ATT AAT AAC TAT TTA TGC TGG TTC CA - #G CAG AAA CCA GGG AAA          480                                                                          Gln Asp Ile Asn Asn Tyr Leu Cys Trp Phe Gl - #n Gln Lys Pro Gly Lys           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - TCT CCT AAG ACC CTG ATC TAT CGT GCA AAC AG - #A CTG GTA GAT GGG GTC          528                                                                          Ser Pro Lys Thr Leu Ile Tyr Arg Ala Asn Ar - #g Leu Val Asp Gly Val           #               175                                                           - CCA TCA AGG TTC AGT GGC AGT GGA TCT GGA CA - #A GAT TAT TCT CTC ACC          576                                                                          Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Gl - #n Asp Tyr Ser Leu Thr           #           190                                                               - ATT AGC AGC CTG GAG TAT GAA GAT ATG GGA AT - #T TAT TAT TGT CTA CAG          624                                                                          Ile Ser Ser Leu Glu Tyr Glu Asp Met Gly Il - #e Tyr Tyr Cys Leu Gln           #       205                                                                   - TAT GAT GAG TTT CCG TAC ACG TTC GGA GGG GG - #G ACC AAG CTG GAA ATA          672                                                                          Tyr Asp Glu Phe Pro Tyr Thr Phe Gly Gly Gl - #y Thr Lys Leu Glu Ile           #   220                                                                       - AAA GGA GGC GGT GGC TCG GGC GGT GGC GGA TC - #G GGT GGC GGC GGC TCT          720                                                                          Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Se - #r Gly Gly Gly Gly Ser           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GAG GTT CAG CTC CAG CAG TCT GGG ACT GTA CT - #G GCA AGG CCT GGG GCT          768                                                                          Glu Val Gln Leu Gln Gln Ser Gly Thr Val Le - #u Ala Arg Pro Gly Ala           #               255                                                           - TCA GTG AAG ATG TCC TGC AAG GCT TCT GGC TA - #C ACC TTT TCC AGC TAC          816                                                                          Ser Val Lys Met Ser Cys Lys Ala Ser Gly Ty - #r Thr Phe Ser Ser Tyr           #           270                                                               - TGG ATG CAC TGG ATA AAA CAG AGG CCT GGA CA - #G GGT CTG GAC TGG ATT          864                                                                          Trp Met His Trp Ile Lys Gln Arg Pro Gly Gl - #n Gly Leu Asp Trp Ile           #       285                                                                   - GTC GCT ATT GAT CCT CGA AAT AGT GAT ACT AT - #T TAC AAC CCG CAA TTC          912                                                                          Val Ala Ile Asp Pro Arg Asn Ser Asp Thr Il - #e Tyr Asn Pro Gln Phe           #   300                                                                       - AAA CAC AAG GCC AAA CTG ACT GCA GTC ACC TC - #C ACC AGC ACT GCC TAC          960                                                                          Lys His Lys Ala Lys Leu Thr Ala Val Thr Se - #r Thr Ser Thr Ala Tyr           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - ATG GAA CTC AAC AGC CTG ACA AAT GAG GAC TC - #T GCG GTC TAT TAC TGT         1008                                                                          Met Glu Leu Asn Ser Leu Thr Asn Glu Asp Se - #r Ala Val Tyr Tyr Cys           #               335                                                           - ACC CCT CTT TAT TAC TTT GAC TCC TGG GGC CA - #A GGC ACC ACT CTC ACA         1056                                                                          Thr Pro Leu Tyr Tyr Phe Asp Ser Trp Gly Gl - #n Gly Thr Thr Leu Thr           #           350                                                               #1074              AC CAT                                                     Val Ser Ser His His His                                                               355                                                                   - (2) INFORMATION FOR SEQ ID NO:45:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 358 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Ala Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala         #           110                                                               - Gln Ala Pro Lys Ser Asp Asp Ile Lys Met Th - #r Gln Ser Pro Ser Ser         #       125                                                                   - Met Tyr Ala Ser Leu Gly Glu Arg Val Thr Ph - #e Thr Cys Lys Ala Ser         #   140                                                                       - Gln Asp Ile Asn Asn Tyr Leu Cys Trp Phe Gl - #n Gln Lys Pro Gly Lys         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Ser Pro Lys Thr Leu Ile Tyr Arg Ala Asn Ar - #g Leu Val Asp Gly Val         #               175                                                           - Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Gl - #n Asp Tyr Ser Leu Thr         #           190                                                               - Ile Ser Ser Leu Glu Tyr Glu Asp Met Gly Il - #e Tyr Tyr Cys Leu Gln         #       205                                                                   - Tyr Asp Glu Phe Pro Tyr Thr Phe Gly Gly Gl - #y Thr Lys Leu Glu Ile         #   220                                                                       - Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Se - #r Gly Gly Gly Gly Ser         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Glu Val Gln Leu Gln Gln Ser Gly Thr Val Le - #u Ala Arg Pro Gly Ala         #               255                                                           - Ser Val Lys Met Ser Cys Lys Ala Ser Gly Ty - #r Thr Phe Ser Ser Tyr         #           270                                                               - Trp Met His Trp Ile Lys Gln Arg Pro Gly Gl - #n Gly Leu Asp Trp Ile         #       285                                                                   - Val Ala Ile Asp Pro Arg Asn Ser Asp Thr Il - #e Tyr Asn Pro Gln Phe         #   300                                                                       - Lys His Lys Ala Lys Leu Thr Ala Val Thr Se - #r Thr Ser Thr Ala Tyr         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Met Glu Leu Asn Ser Leu Thr Asn Glu Asp Se - #r Ala Val Tyr Tyr Cys         #               335                                                           - Thr Pro Leu Tyr Tyr Phe Asp Ser Trp Gly Gl - #n Gly Thr Thr Leu Thr         #           350                                                               - Val Ser Ser His His His                                                             355                                                                   - (2) INFORMATION FOR SEQ ID NO:46:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1086 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1083                                               #/note= "MetSerOncAngsFv"MATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT GTT GTT GCT TG - #T GAA AAT GGC TTA CCT          288                                                                          Lys Lys Ser Thr Asn Lys Phe Val Val Ala Cy - #s Glu Asn Gly Leu Pro           #                 95                                                          - GTC CAC TTG GAT CAG TCA ATT TTC CGT CGT CC - #G GCC AAG AAA CTG AAC          336                                                                          Val His Leu Asp Gln Ser Ile Phe Arg Arg Pr - #o Ala Lys Lys Leu Asn           #           110                                                               - GAC GCT CAG GCG CCG AAG AGT GAT GAC ATC AA - #G ATG ACC CAG TCT CCA          384                                                                          Asp Ala Gln Ala Pro Lys Ser Asp Asp Ile Ly - #s Met Thr Gln Ser Pro           #       125                                                                   - TCT TCC ATG TAT GCA TCT CTA GGA GAG AGA GT - #C ACT TTC ACT TGC AAG          432                                                                          Ser Ser Met Tyr Ala Ser Leu Gly Glu Arg Va - #l Thr Phe Thr Cys Lys           #   140                                                                       - GCG AGT CAG GAC ATT AAT AAC TAT TTA TGC TG - #G TTC CAG CAG AAA CCA          480                                                                          Ala Ser Gln Asp Ile Asn Asn Tyr Leu Cys Tr - #p Phe Gln Gln Lys Pro           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - GGG AAA TCT CCT AAG ACC CTG ATC TAT CGT GC - #A AAC AGA CTG GTA GAT          528                                                                          Gly Lys Ser Pro Lys Thr Leu Ile Tyr Arg Al - #a Asn Arg Leu Val Asp           #               175                                                           - GGG GTC CCA TCA AGG TTC AGT GGC AGT GGA TC - #T GGA CAA GAT TAT TCT          576                                                                          Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Se - #r Gly Gln Asp Tyr Ser           #           190                                                               - CTC ACC ATT AGC AGC CTG GAG TAT GAA GAT AT - #G GGA ATT TAT TAT TGT          624                                                                          Leu Thr Ile Ser Ser Leu Glu Tyr Glu Asp Me - #t Gly Ile Tyr Tyr Cys           #       205                                                                   - CTA CAG TAT GAT GAG TTT CCG TAC ACG TTC GG - #A GGG GGG ACC AAG CTG          672                                                                          Leu Gln Tyr Asp Glu Phe Pro Tyr Thr Phe Gl - #y Gly Gly Thr Lys Leu           #   220                                                                       - GAA ATA AAA GGA GGC GGT GGC TCG GGC GGT GG - #C GGA TCG GGT GGC GGC          720                                                                          Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gl - #y Gly Ser Gly Gly Gly           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GGC TCT GAG GTT CAG CTC CAG CAG TCT GGG AC - #T GTA CTG GCA AGG CCT          768                                                                          Gly Ser Glu Val Gln Leu Gln Gln Ser Gly Th - #r Val Leu Ala Arg Pro           #               255                                                           - GGG GCT TCA GTG AAG ATG TCC TGC AAG GCT TC - #T GGC TAC ACC TTT TCC          816                                                                          Gly Ala Ser Val Lys Met Ser Cys Lys Ala Se - #r Gly Tyr Thr Phe Ser           #           270                                                               - AGC TAC TGG ATG CAC TGG ATA AAA CAG AGG CC - #T GGA CAG GGT CTG GAC          864                                                                          Ser Tyr Trp Met His Trp Ile Lys Gln Arg Pr - #o Gly Gln Gly Leu Asp           #       285                                                                   - TGG ATT GTC GCT ATT GAT CCT CGA AAT AGT GA - #T ACT ATT TAC AAC CCG          912                                                                          Trp Ile Val Ala Ile Asp Pro Arg Asn Ser As - #p Thr Ile Tyr Asn Pro           #   300                                                                       - CAA TTC AAA CAC AAG GCC AAA CTG ACT GCA GT - #C ACC TCC ACC AGC ACT          960                                                                          Gln Phe Lys His Lys Ala Lys Leu Thr Ala Va - #l Thr Ser Thr Ser Thr           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - GCC TAC ATG GAA CTC AAC AGC CTG ACA AAT GA - #G GAC TCT GCG GTC TAT         1008                                                                          Ala Tyr Met Glu Leu Asn Ser Leu Thr Asn Gl - #u Asp Ser Ala Val Tyr           #               335                                                           - TAC TGT ACC CCT CTT TAT TAC TTT GAC TCC TG - #G GGC CAA GGC ACC ACT         1056                                                                          Tyr Cys Thr Pro Leu Tyr Tyr Phe Asp Ser Tr - #p Gly Gln Gly Thr Thr           #           350                                                               #         1086     CA CAT CAC CAT TAGTAG                                      Leu Thr Val Ser Ser His His His                                               #       360                                                                   - (2) INFORMATION FOR SEQ ID NO:47:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 360 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Val Val Ala Cy - #s Glu Asn Gly Leu Pro         #                 95                                                          - Val His Leu Asp Gln Ser Ile Phe Arg Arg Pr - #o Ala Lys Lys Leu Asn         #           110                                                               - Asp Ala Gln Ala Pro Lys Ser Asp Asp Ile Ly - #s Met Thr Gln Ser Pro         #       125                                                                   - Ser Ser Met Tyr Ala Ser Leu Gly Glu Arg Va - #l Thr Phe Thr Cys Lys         #   140                                                                       - Ala Ser Gln Asp Ile Asn Asn Tyr Leu Cys Tr - #p Phe Gln Gln Lys Pro         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Gly Lys Ser Pro Lys Thr Leu Ile Tyr Arg Al - #a Asn Arg Leu Val Asp         #               175                                                           - Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Se - #r Gly Gln Asp Tyr Ser         #           190                                                               - Leu Thr Ile Ser Ser Leu Glu Tyr Glu Asp Me - #t Gly Ile Tyr Tyr Cys         #       205                                                                   - Leu Gln Tyr Asp Glu Phe Pro Tyr Thr Phe Gl - #y Gly Gly Thr Lys Leu         #   220                                                                       - Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gl - #y Gly Ser Gly Gly Gly         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Gly Ser Glu Val Gln Leu Gln Gln Ser Gly Th - #r Val Leu Ala Arg Pro         #               255                                                           - Gly Ala Ser Val Lys Met Ser Cys Lys Ala Se - #r Gly Tyr Thr Phe Ser         #           270                                                               - Ser Tyr Trp Met His Trp Ile Lys Gln Arg Pr - #o Gly Gln Gly Leu Asp         #       285                                                                   - Trp Ile Val Ala Ile Asp Pro Arg Asn Ser As - #p Thr Ile Tyr Asn Pro         #   300                                                                       - Gln Phe Lys His Lys Ala Lys Leu Thr Ala Va - #l Thr Ser Thr Ser Thr         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Ala Tyr Met Glu Leu Asn Ser Leu Thr Asn Gl - #u Asp Ser Ala Val Tyr         #               335                                                           - Tyr Cys Thr Pro Leu Tyr Tyr Phe Asp Ser Tr - #p Gly Gln Gly Thr Thr         #           350                                                               - Leu Thr Val Ser Ser His His His                                             #       360                                                                   - (2) INFORMATION FOR SEQ ID NO:48:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1065 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1065                                               #/note= "sFvOncMetSer"FORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                - GAC ATC AAG ATG ACC CAG TCT CCA TCT TCC AT - #G TAT GCA TCT CTA GGA           48                                                                          Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Me - #t Tyr Ala Ser Leu Gly           #                 15                                                          - GAG AGA GTC ACT TTC ACT TGC AAG GCG AGT CA - #G GAC ATT AAT AAC TAT           96                                                                          Glu Arg Val Thr Phe Thr Cys Lys Ala Ser Gl - #n Asp Ile Asn Asn Tyr           #             30                                                              - TTA TGC TGG TTC CAG CAG AAA CCA GGG AAA TC - #T CCT AAG ACC CTG ATC          144                                                                          Leu Cys Trp Phe Gln Gln Lys Pro Gly Lys Se - #r Pro Lys Thr Leu Ile           #         45                                                                  - TAT CGT GCA AAC AGA CTG GTA GAT GGG GTC CC - #A TCA AGG TTC AGT GGC          192                                                                          Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pr - #o Ser Arg Phe Ser Gly           #     60                                                                      - AGT GGA TCT GGA CAA GAT TAT TCT CTC ACC AT - #T AGC AGC CTG GAG TAT          240                                                                          Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Il - #e Ser Ser Leu Glu Tyr           # 80                                                                          - GAA GAT ATG GGA ATT TAT TAT TGT CTA CAG TA - #T GAT GAG TTT CCG TAC          288                                                                          Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Ty - #r Asp Glu Phe Pro Tyr           #                 95                                                          - ACG TTC GGA GGG GGG ACC AAG CTG GAA ATA AA - #A GGA GGC GGT GGC TCG          336                                                                          Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Ly - #s Gly Gly Gly Gly Ser           #           110                                                               - GGC GGT GGC GGA TCG GGT GGC GGC GGC TCT GA - #G GTT CAG CTC CAG CAG          384                                                                          Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #u Val Gln Leu Gln Gln           #       125                                                                   - TCT GGG ACT GTA CTG GCA AGG CCT GGG GCT TC - #A GTG AAG ATG TCC TGC          432                                                                          Ser Gly Thr Val Leu Ala Arg Pro Gly Ala Se - #r Val Lys Met Ser Cys           #   140                                                                       - AAG GCT TCT GGC TAC ACC TTT TCC AGC TAC TG - #G ATG CAC TGG ATA AAA          480                                                                          Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Tr - #p Met His Trp Ile Lys           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - CAG AGG CCT GGA CAG GGT CTG GAC TGG ATT GT - #C GCT ATT GAT CCT CGA          528                                                                          Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile Va - #l Ala Ile Asp Pro Arg           #               175                                                           - AAT AGT GAT ACT ATT TAC AAC CCG CAA TTC AA - #A CAC AAG GCC AAA CTG          576                                                                          Asn Ser Asp Thr Ile Tyr Asn Pro Gln Phe Ly - #s His Lys Ala Lys Leu           #           190                                                               - ACT GCA GTC ACC TCC ACC AGC ACT GCC TAC AT - #G GAA CTC AAC AGC CTG          624                                                                          Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr Me - #t Glu Leu Asn Ser Leu           #       205                                                                   - ACA AAT GAG GAC TCT GCG GTC TAT TAC TGT AC - #C CCT CTT TAT TAC TTT          672                                                                          Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys Th - #r Pro Leu Tyr Tyr Phe           #   220                                                                       - GAC TCC TGG GGC CAA GGC ACC ACT CTC ACA GT - #C TCC TCA GCC AAG AAA          720                                                                          Asp Ser Trp Gly Gln Gly Thr Thr Leu Thr Va - #l Ser Ser Ala Lys Lys           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - CTG AAC GAC GCT CAG GCG CCG AAG AGT GAT AT - #G TCA GAT TGG CTT ACA          768                                                                          Leu Asn Asp Ala Gln Ala Pro Lys Ser Asp Me - #t Ser Asp Trp Leu Thr           #               255                                                           - TTT CAG AAA AAA CAC ATC ACA AAC ACA AGG GA - #T GTT GAT TGT GAT AAT          816                                                                          Phe Gln Lys Lys His Ile Thr Asn Thr Arg As - #p Val Asp Cys Asp Asn           #           270                                                               - ATC ATG TCA ACA AAC TTG TTC CAC TGC AAG GA - #C AAG AAC ACT TTT ATC          864                                                                          Ile Met Ser Thr Asn Leu Phe His Cys Lys As - #p Lys Asn Thr Phe Ile           #       285                                                                   - TAT TCA CGT CCT GAG CCA GTG AAG GCC ATC TG - #T AAA GGA ATT ATA GCC          912                                                                          Tyr Ser Arg Pro Glu Pro Val Lys Ala Ile Cy - #s Lys Gly Ile Ile Ala           #   300                                                                       - TCC AAA AAT GTG TTA ACT ACC TCT GAG TTT TA - #T CTC TCT GAT TGC AAT          960                                                                          Ser Lys Asn Val Leu Thr Thr Ser Glu Phe Ty - #r Leu Ser Asp Cys Asn           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - GTA ACA AGC AGG CCT TGC AAG TAT AAA TTA AA - #G AAA TCA ACT AAT AAA         1008                                                                          Val Thr Ser Arg Pro Cys Lys Tyr Lys Leu Ly - #s Lys Ser Thr Asn Lys           #               335                                                           - TTT TGT GTA ACT TGT GAA AAT CAG GCA CCA GT - #T CAT TTT GTT GGA GTT         1056                                                                          Phe Cys Val Thr Cys Glu Asn Gln Ala Pro Va - #l His Phe Val Gly Val           #           350                                                               #       1065                                                                  Gly Ser Cys                                                                           355                                                                   - (2) INFORMATION FOR SEQ ID NO:49:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 355 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                - Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Me - #t Tyr Ala Ser Leu Gly         #                 15                                                          - Glu Arg Val Thr Phe Thr Cys Lys Ala Ser Gl - #n Asp Ile Asn Asn Tyr         #             30                                                              - Leu Cys Trp Phe Gln Gln Lys Pro Gly Lys Se - #r Pro Lys Thr Leu Ile         #         45                                                                  - Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pr - #o Ser Arg Phe Ser Gly         #     60                                                                      - Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Il - #e Ser Ser Leu Glu Tyr         # 80                                                                          - Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Ty - #r Asp Glu Phe Pro Tyr         #                 95                                                          - Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Ly - #s Gly Gly Gly Gly Ser         #           110                                                               - Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #u Val Gln Leu Gln Gln         #       125                                                                   - Ser Gly Thr Val Leu Ala Arg Pro Gly Ala Se - #r Val Lys Met Ser Cys         #   140                                                                       - Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Tr - #p Met His Trp Ile Lys         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile Va - #l Ala Ile Asp Pro Arg         #               175                                                           - Asn Ser Asp Thr Ile Tyr Asn Pro Gln Phe Ly - #s His Lys Ala Lys Leu         #           190                                                               - Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr Me - #t Glu Leu Asn Ser Leu         #       205                                                                   - Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys Th - #r Pro Leu Tyr Tyr Phe         #   220                                                                       - Asp Ser Trp Gly Gln Gly Thr Thr Leu Thr Va - #l Ser Ser Ala Lys Lys         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Leu Asn Asp Ala Gln Ala Pro Lys Ser Asp Me - #t Ser Asp Trp Leu Thr         #               255                                                           - Phe Gln Lys Lys His Ile Thr Asn Thr Arg As - #p Val Asp Cys Asp Asn         #           270                                                               - Ile Met Ser Thr Asn Leu Phe His Cys Lys As - #p Lys Asn Thr Phe Ile         #       285                                                                   - Tyr Ser Arg Pro Glu Pro Val Lys Ala Ile Cy - #s Lys Gly Ile Ile Ala         #   300                                                                       - Ser Lys Asn Val Leu Thr Thr Ser Glu Phe Ty - #r Leu Ser Asp Cys Asn         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Val Thr Ser Arg Pro Cys Lys Tyr Lys Leu Ly - #s Lys Ser Thr Asn Lys         #               335                                                           - Phe Cys Val Thr Cys Glu Asn Gln Ala Pro Va - #l His Phe Val Gly Val         #           350                                                               - Gly Ser Cys                                                                         355                                                                   - (2) INFORMATION FOR SEQ ID NO:50:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1074 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1074                                               #/note= "MetGluOncFBE6"ORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                - ATG GAG GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Glu Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          - GTT CAT TTT GTT GGA GTT GGA TCT TGT GCC AA - #G AAA CTG AAC GAC GCT          336                                                                          Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala           #           110                                                               - CAG GCG CCG AAG AGT GAT GAC ATC AAG ATG AC - #C CAG TCT CCA TCT TCC          384                                                                          Gln Ala Pro Lys Ser Asp Asp Ile Lys Met Th - #r Gln Ser Pro Ser Ser           #       125                                                                   - ATG TAT GCA TCT CTA GGA GAG AGA GTC ACT TT - #C ACT TGC AAG GCG AGT          432                                                                          Met Tyr Ala Ser Leu Gly Glu Arg Val Thr Ph - #e Thr Cys Lys Ala Ser           #   140                                                                       - CAG GAC ATT AAT AAC TAT TTA TGC TGG TTC CA - #G CAG AAA CCA GGG AAA          480                                                                          Gln Asp Ile Asn Asn Tyr Leu Cys Trp Phe Gl - #n Gln Lys Pro Gly Lys           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - TCT CCT AAG ACC CTG ATC TAT CGT GCA AAC AG - #A CTG GTA GAT GGG GTC          528                                                                          Ser Pro Lys Thr Leu Ile Tyr Arg Ala Asn Ar - #g Leu Val Asp Gly Val           #               175                                                           - CCA TCA AGG TTC AGT GGC AGT GGA TCT GGA CA - #A GAT TAT TCT CTC ACC          576                                                                          Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Gl - #n Asp Tyr Ser Leu Thr           #           190                                                               - ATT AGC AGC CTG GAG TAT GAA GAT ATG GGA AT - #T TAT TAT TGT CTA CAG          624                                                                          Ile Ser Ser Leu Glu Tyr Glu Asp Met Gly Il - #e Tyr Tyr Cys Leu Gln           #       205                                                                   - TAT GAT GAG TTT CCG TAC ACG TTC GGA GGG GG - #G ACC AAG CTG GAA ATA          672                                                                          Tyr Asp Glu Phe Pro Tyr Thr Phe Gly Gly Gl - #y Thr Lys Leu Glu Ile           #   220                                                                       - AAA GGA GGC GGT GGC TCG GGC GGT GGC GGA TC - #G GGT GGC GGC GGC TCT          720                                                                          Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Se - #r Gly Gly Gly Gly Ser           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GAG GTT CAG CTC CAG CAG TCT GGG ACT GTA CT - #G GCA AGG CCT GGG GCT          768                                                                          Glu Val Gln Leu Gln Gln Ser Gly Thr Val Le - #u Ala Arg Pro Gly Ala           #               255                                                           - TCA GTG AAG ATG TCC TGC AAG GCT TCT GGC TA - #C ACC TTT TCC AGC TAC          816                                                                          Ser Val Lys Met Ser Cys Lys Ala Ser Gly Ty - #r Thr Phe Ser Ser Tyr           #           270                                                               - TGG ATG CAC TGG ATA AAA CAG AGG CCT GGA CA - #G GGT CTG GAC TGG ATT          864                                                                          Trp Met His Trp Ile Lys Gln Arg Pro Gly Gl - #n Gly Leu Asp Trp Ile           #       285                                                                   - GTC GCT ATT GAT CCT CGA AAT AGT GAT ACT AT - #T TAC AAC CCG CAA TTC          912                                                                          Val Ala Ile Asp Pro Arg Asn Ser Asp Thr Il - #e Tyr Asn Pro Gln Phe           #   300                                                                       - AAA CAC AAG GCC AAA CTG ACT GCA GTC ACC TC - #C ACC AGC ACT GCC TAC          960                                                                          Lys His Lys Ala Lys Leu Thr Ala Val Thr Se - #r Thr Ser Thr Ala Tyr           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - ATG GAA CTC AAC AGC CTG ACA AAT GAG GAC TC - #T GCG GTC TAT TAC TGT         1008                                                                          Met Glu Leu Asn Ser Leu Thr Asn Glu Asp Se - #r Ala Val Tyr Tyr Cys           #               335                                                           - ACC CCT CTT TAT TAC TTT GAC TCC TGG GGC CA - #A GGC ACC ACT CTC ACA         1056                                                                          Thr Pro Leu Tyr Tyr Phe Asp Ser Trp Gly Gl - #n Gly Thr Thr Leu Thr           #           350                                                               #1074              AC CAT                                                     Val Ser Ser His His His                                                               355                                                                   - (2) INFORMATION FOR SEQ ID NO:51:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 358 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                - Met Glu Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala         #           110                                                               - Gln Ala Pro Lys Ser Asp Asp Ile Lys Met Th - #r Gln Ser Pro Ser Ser         #       125                                                                   - Met Tyr Ala Ser Leu Gly Glu Arg Val Thr Ph - #e Thr Cys Lys Ala Ser         #   140                                                                       - Gln Asp Ile Asn Asn Tyr Leu Cys Trp Phe Gl - #n Gln Lys Pro Gly Lys         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Ser Pro Lys Thr Leu Ile Tyr Arg Ala Asn Ar - #g Leu Val Asp Gly Val         #               175                                                           - Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Gl - #n Asp Tyr Ser Leu Thr         #           190                                                               - Ile Ser Ser Leu Glu Tyr Glu Asp Met Gly Il - #e Tyr Tyr Cys Leu Gln         #       205                                                                   - Tyr Asp Glu Phe Pro Tyr Thr Phe Gly Gly Gl - #y Thr Lys Leu Glu Ile         #   220                                                                       - Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Se - #r Gly Gly Gly Gly Ser         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Glu Val Gln Leu Gln Gln Ser Gly Thr Val Le - #u Ala Arg Pro Gly Ala         #               255                                                           - Ser Val Lys Met Ser Cys Lys Ala Ser Gly Ty - #r Thr Phe Ser Ser Tyr         #           270                                                               - Trp Met His Trp Ile Lys Gln Arg Pro Gly Gl - #n Gly Leu Asp Trp Ile         #       285                                                                   - Val Ala Ile Asp Pro Arg Asn Ser Asp Thr Il - #e Tyr Asn Pro Gln Phe         #   300                                                                       - Lys His Lys Ala Lys Leu Thr Ala Val Thr Se - #r Thr Ser Thr Ala Tyr         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Met Glu Leu Asn Ser Leu Thr Asn Glu Asp Se - #r Ala Val Tyr Tyr Cys         #               335                                                           - Thr Pro Leu Tyr Tyr Phe Asp Ser Trp Gly Gl - #n Gly Thr Thr Leu Thr         #           350                                                               - Val Ser Ser His His His                                                             355                                                                   - (2) INFORMATION FOR SEQ ID NO:52:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1095 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1095                                               #/note= "MetSerOncA87FBMOC31"ON:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT GCT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Ala Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          - GTT CAT TTT GTT GGA GTT GGA TCT TGT GCC AA - #G AAA CTG AAC GAC GCT          336                                                                          Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala           #           110                                                               - CAG GCG CCG AAG AGT GAT CAG GTG AAG CTG CA - #G CAG TCA GGA CCT GAG          384                                                                          Gln Ala Pro Lys Ser Asp Gln Val Lys Leu Gl - #n Gln Ser Gly Pro Glu           #       125                                                                   - CTG AAG AAG CCT GGA GAG ACA GTC AAG ATC TC - #C TGC AAG GCT TCT GGG          432                                                                          Leu Lys Lys Pro Gly Glu Thr Val Lys Ile Se - #r Cys Lys Ala Ser Gly           #   140                                                                       - TAC ACC TTC ACA AAC TAT GGA ATG AAC TGG GT - #G AAG CAG GCT CCA GGA          480                                                                          Tyr Thr Phe Thr Asn Tyr Gly Met Asn Trp Va - #l Lys Gln Ala Pro Gly           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - AAG GGT TTA AAG TGG ATG GGC TGG ATA AAC AC - #C TAC ACT GGA GAG TCA          528                                                                          Lys Gly Leu Lys Trp Met Gly Trp Ile Asn Th - #r Tyr Thr Gly Glu Ser           #               175                                                           - ACA TAT GCT GAT GAC TTC AAG GGA CGG TTT GC - #C TTT TCT CTA GAA ACC          576                                                                          Thr Tyr Ala Asp Asp Phe Lys Gly Arg Phe Al - #a Phe Ser Leu Glu Thr           #           190                                                               - TCT GCC AGC GCT GCC TAT TTG CAG ATC AAC AA - #C CTC AAA AAT GAG GAC          624                                                                          Ser Ala Ser Ala Ala Tyr Leu Gln Ile Asn As - #n Leu Lys Asn Glu Asp           #       205                                                                   - ACG GCT ACA TAT TTC TGT GCA AGA TTC GCT AT - #T AAG GGG GAC TAC TGG          672                                                                          Thr Ala Thr Tyr Phe Cys Ala Arg Phe Ala Il - #e Lys Gly Asp Tyr Trp           #   220                                                                       - GGC CAA GGG ACC ACG GTC ACC GTC TCC TCA GG - #T GGA GGC GGT TCA GGC          720                                                                          Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gl - #y Gly Gly Gly Ser Gly           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GGA GGT GGC TCT GGC GGT GGC GGA TCG GAC AT - #T GTG CTA ACC CAG TCT          768                                                                          Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Il - #e Val Leu Thr Gln Ser           #               255                                                           - CCA TTC TCC AAT CCA GTC ACT CTT GGA ACA TC - #A GCT TCC ATC TCC TGC          816                                                                          Pro Phe Ser Asn Pro Val Thr Leu Gly Thr Se - #r Ala Ser Ile Ser Cys           #           270                                                               - AGG TCT ACT AAG AGT CTC CTA CAT AGT AAT GG - #C ATC ACT TAT TTG TAT          864                                                                          Arg Ser Thr Lys Ser Leu Leu His Ser Asn Gl - #y Ile Thr Tyr Leu Tyr           #       285                                                                   - TGG TAT CTG CAG AAG CCA GGC CAG TCT CCT CA - #G CTC CTG ATT TAT CAG          912                                                                          Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gl - #n Leu Leu Ile Tyr Gln           #   300                                                                       - ATG TCC AAC CTT GCC TCA GGA GTC CCA GAC AG - #G TTC AGT AGC AGT GGG          960                                                                          Met Ser Asn Leu Ala Ser Gly Val Pro Asp Ar - #g Phe Ser Ser Ser Gly           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - TCA GGA ACT GAT TTC ACA CTG AGA ATC AGC AG - #A GTG GAG GCT GAG GAT         1008                                                                          Ser Gly Thr Asp Phe Thr Leu Arg Ile Ser Ar - #g Val Glu Ala Glu Asp           #               335                                                           - GTG GGT GTT TAT TAC TGT GCT CAA AAT CTA GA - #A ATT CCT CGG ACG TTC         1056                                                                          Val Gly Val Tyr Tyr Cys Ala Gln Asn Leu Gl - #u Ile Pro Arg Thr Phe           #           350                                                               #   1095A GGC ACC AAG CTG GAA ATC AAA CGG GC - #G GCC GCA                     Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Al - #a Ala Ala                       #       365                                                                   - (2) INFORMATION FOR SEQ ID NO:53:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 365 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Ala Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala         #           110                                                               - Gln Ala Pro Lys Ser Asp Gln Val Lys Leu Gl - #n Gln Ser Gly Pro Glu         #       125                                                                   - Leu Lys Lys Pro Gly Glu Thr Val Lys Ile Se - #r Cys Lys Ala Ser Gly         #   140                                                                       - Tyr Thr Phe Thr Asn Tyr Gly Met Asn Trp Va - #l Lys Gln Ala Pro Gly         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Lys Gly Leu Lys Trp Met Gly Trp Ile Asn Th - #r Tyr Thr Gly Glu Ser         #               175                                                           - Thr Tyr Ala Asp Asp Phe Lys Gly Arg Phe Al - #a Phe Ser Leu Glu Thr         #           190                                                               - Ser Ala Ser Ala Ala Tyr Leu Gln Ile Asn As - #n Leu Lys Asn Glu Asp         #       205                                                                   - Thr Ala Thr Tyr Phe Cys Ala Arg Phe Ala Il - #e Lys Gly Asp Tyr Trp         #   220                                                                       - Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gl - #y Gly Gly Gly Ser Gly         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Il - #e Val Leu Thr Gln Ser         #               255                                                           - Pro Phe Ser Asn Pro Val Thr Leu Gly Thr Se - #r Ala Ser Ile Ser Cys         #           270                                                               - Arg Ser Thr Lys Ser Leu Leu His Ser Asn Gl - #y Ile Thr Tyr Leu Tyr         #       285                                                                   - Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gl - #n Leu Leu Ile Tyr Gln         #   300                                                                       - Met Ser Asn Leu Ala Ser Gly Val Pro Asp Ar - #g Phe Ser Ser Ser Gly         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Ser Gly Thr Asp Phe Thr Leu Arg Ile Ser Ar - #g Val Glu Ala Glu Asp         #               335                                                           - Val Gly Val Tyr Tyr Cys Ala Gln Asn Leu Gl - #u Ile Pro Arg Thr Phe         #           350                                                               - Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Al - #a Ala Ala                     #       365                                                                   - (2) INFORMATION FOR SEQ ID NO:54:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1098 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1098                                               #/note= "MOC31FBMetSerOnc"ATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                - GGT CAG GTG AAG CTG CAG CAG TCA GGA CCT GA - #G CTG AAG AAG CCT GGA           48                                                                          Gly Gln Val Lys Leu Gln Gln Ser Gly Pro Gl - #u Leu Lys Lys Pro Gly           #                 15                                                          - GAG ACA GTC AAG ATC TCC TGC AAG GCT TCT GG - #G TAC ACC TTC ACA AAC           96                                                                          Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gl - #y Tyr Thr Phe Thr Asn           #             30                                                              - TAT GGA ATG AAC TGG GTG AAG CAG GCT CCA GG - #A AAG GGT TTA AAG TGG          144                                                                          Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gl - #y Lys Gly Leu Lys Trp           #         45                                                                  - ATG GGC TGG ATA AAC ACC TAC ACT GGA GAG TC - #A ACA TAT GCT GAT GAC          192                                                                          Met Gly Trp Ile Asn Thr Tyr Thr Gly Glu Se - #r Thr Tyr Ala Asp Asp           #     60                                                                      - TTC AAG GGA CGG TTT GCC TTT TCT CTA GAA AC - #C TCT GCC AGC GCT GCC          240                                                                          Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Th - #r Ser Ala Ser Ala Ala           # 80                                                                          - TAT TTG CAG ATC AAC AAC CTC AAA AAT GAG GA - #C ACG GCT ACA TAT TTC          288                                                                          Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu As - #p Thr Ala Thr Tyr Phe           #                 95                                                          - TGT GCA AGA TTC GCT ATT AAG GGG GAC TAC TG - #G GGC CAA GGG ACC ACG          336                                                                          Cys Ala Arg Phe Ala Ile Lys Gly Asp Tyr Tr - #p Gly Gln Gly Thr Thr           #           110                                                               - GTC ACC GTC TCC TCA GGT GGA GGC GGT TCA GG - #C GGA GGT GGC TCT GGC          384                                                                          Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gl - #y Gly Gly Gly Ser Gly           #       125                                                                   - GGT GGC GGA TCG GAC ATT GTG CTA ACC CAG TC - #T CCA TTC TCC AAT CCA          432                                                                          Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Se - #r Pro Phe Ser Asn Pro           #   140                                                                       - GTC ACT CTT GGA ACA TCA GCT TCC ATC TCC TG - #C AGG TCT ACT AAG AGT          480                                                                          Val Thr Leu Gly Thr Ser Ala Ser Ile Ser Cy - #s Arg Ser Thr Lys Ser           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - CTC CTA CAT AGT AAT GGC ATC ACT TAT TTG TA - #T TGG TAT CTG CAG AAG          528                                                                          Leu Leu His Ser Asn Gly Ile Thr Tyr Leu Ty - #r Trp Tyr Leu Gln Lys           #               175                                                           - CCA GGC CAG TCT CCT CAG CTC CTG ATT TAT CA - #G ATG TCC AAC CTT GCC          576                                                                          Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Gl - #n Met Ser Asn Leu Ala           #           190                                                               - TCA GGA GTC CCA GAC AGG TTC AGT AGC AGT GG - #G TCA GGA ACT GAT TTC          624                                                                          Ser Gly Val Pro Asp Arg Phe Ser Ser Ser Gl - #y Ser Gly Thr Asp Phe           #       205                                                                   - ACA CTG AGA ATC AGC AGA GTG GAG GCT GAG GA - #T GTG GGT GTT TAT TAC          672                                                                          Thr Leu Arg Ile Ser Arg Val Glu Ala Glu As - #p Val Gly Val Tyr Tyr           #   220                                                                       - TGT GCT CAA AAT CTA GAA ATT CCT CGG ACG TT - #C GGT GGA GGC ACC AAG          720                                                                          Cys Ala Gln Asn Leu Glu Ile Pro Arg Thr Ph - #e Gly Gly Gly Thr Lys           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - CTG GAA ATC AAA CGG GCG GCC GCA GCC AAG AA - #A CTG AAC GAC GCT CAG          768                                                                          Leu Glu Ile Lys Arg Ala Ala Ala Ala Lys Ly - #s Leu Asn Asp Ala Gln           #               255                                                           - GCG CCG AAG AGT GAT ATG TCA GAT TGG CTT AC - #A TTT CAG AAA AAA CAC          816                                                                          Ala Pro Lys Ser Asp Met Ser Asp Trp Leu Th - #r Phe Gln Lys Lys His           #           270                                                               - ATC ACA AAC ACA AGG GAT GTT GAT TGT GAT AA - #T ATC ATG TCA ACA AAC          864                                                                          Ile Thr Asn Thr Arg Asp Val Asp Cys Asp As - #n Ile Met Ser Thr Asn           #       285                                                                   - TTG TTC CAC TGC AAG GAC AAG AAC ACT TTT AT - #C TAT TCA CGT CCT GAG          912                                                                          Leu Phe His Cys Lys Asp Lys Asn Thr Phe Il - #e Tyr Ser Arg Pro Glu           #   300                                                                       - CCA GTG AAG GCC ATC TGT AAA GGA ATT ATA GC - #C TCC AAA AAT GTG TTA          960                                                                          Pro Val Lys Ala Ile Cys Lys Gly Ile Ile Al - #a Ser Lys Asn Val Leu           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - ACT ACC TCT GAG TTT TAT CTC TCT GAT TGC AA - #T GTA ACA AGC AGG CCT         1008                                                                          Thr Thr Ser Glu Phe Tyr Leu Ser Asp Cys As - #n Val Thr Ser Arg Pro           #               335                                                           - TGC AAG TAT AAA TTA AAG AAA TCA ACT AAT AA - #A TTT TGT GTA ACT TGT         1056                                                                          Cys Lys Tyr Lys Leu Lys Lys Ser Thr Asn Ly - #s Phe Cys Val Thr Cys           #           350                                                               - GAA AAT CAG GCA CCA GTT CAT TTT GTT GGA GT - #T GGA TCT TGT                 #1098                                                                         Glu Asn Gln Ala Pro Val His Phe Val Gly Va - #l Gly Ser Cys                   #       365                                                                   - (2) INFORMATION FOR SEQ ID NO:55:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 366 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                - Gly Gln Val Lys Leu Gln Gln Ser Gly Pro Gl - #u Leu Lys Lys Pro Gly         #                 15                                                          - Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gl - #y Tyr Thr Phe Thr Asn         #             30                                                              - Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gl - #y Lys Gly Leu Lys Trp         #         45                                                                  - Met Gly Trp Ile Asn Thr Tyr Thr Gly Glu Se - #r Thr Tyr Ala Asp Asp         #     60                                                                      - Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Th - #r Ser Ala Ser Ala Ala         # 80                                                                          - Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu As - #p Thr Ala Thr Tyr Phe         #                 95                                                          - Cys Ala Arg Phe Ala Ile Lys Gly Asp Tyr Tr - #p Gly Gln Gly Thr Thr         #           110                                                               - Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gl - #y Gly Gly Gly Ser Gly         #       125                                                                   - Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Se - #r Pro Phe Ser Asn Pro         #   140                                                                       - Val Thr Leu Gly Thr Ser Ala Ser Ile Ser Cy - #s Arg Ser Thr Lys Ser         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Leu Leu His Ser Asn Gly Ile Thr Tyr Leu Ty - #r Trp Tyr Leu Gln Lys         #               175                                                           - Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Gl - #n Met Ser Asn Leu Ala         #           190                                                               - Ser Gly Val Pro Asp Arg Phe Ser Ser Ser Gl - #y Ser Gly Thr Asp Phe         #       205                                                                   - Thr Leu Arg Ile Ser Arg Val Glu Ala Glu As - #p Val Gly Val Tyr Tyr         #   220                                                                       - Cys Ala Gln Asn Leu Glu Ile Pro Arg Thr Ph - #e Gly Gly Gly Thr Lys         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Leu Glu Ile Lys Arg Ala Ala Ala Ala Lys Ly - #s Leu Asn Asp Ala Gln         #               255                                                           - Ala Pro Lys Ser Asp Met Ser Asp Trp Leu Th - #r Phe Gln Lys Lys His         #           270                                                               - Ile Thr Asn Thr Arg Asp Val Asp Cys Asp As - #n Ile Met Ser Thr Asn         #       285                                                                   - Leu Phe His Cys Lys Asp Lys Asn Thr Phe Il - #e Tyr Ser Arg Pro Glu         #   300                                                                       - Pro Val Lys Ala Ile Cys Lys Gly Ile Ile Al - #a Ser Lys Asn Val Leu         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Thr Thr Ser Glu Phe Tyr Leu Ser Asp Cys As - #n Val Thr Ser Arg Pro         #               335                                                           - Cys Lys Tyr Lys Leu Lys Lys Ser Thr Asn Ly - #s Phe Cys Val Thr Cys         #           350                                                               - Glu Asn Gln Ala Pro Val His Phe Val Gly Va - #l Gly Ser Cys                 #       365                                                                   - (2) INFORMATION FOR SEQ ID NO:56:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 1065 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1065                                               #/note= "MetSerOncFBMOC161"TION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          - GTT CAT TTT GTT GGA GTT GGA TCT TGT GCC AA - #G AAA CTG AAC GAC GCT          336                                                                          Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala           #           110                                                               - CAG GCG CCG AAG AGT GAT CAG GTC CAA CTG CA - #G CAG TCA GGA ACT GAG          384                                                                          Gln Ala Pro Lys Ser Asp Gln Val Gln Leu Gl - #n Gln Ser Gly Thr Glu           #       125                                                                   - CTG ATA AGG CCT GGG ACT TCA GTG AAG ATA TC - #C TGT AAG GCT TCT GGA          432                                                                          Leu Ile Arg Pro Gly Thr Ser Val Lys Ile Se - #r Cys Lys Ala Ser Gly           #   140                                                                       - TAC GCC TTC ACT GAC TAC TGG CTA GGT TGG GT - #A AAA CAC AGG CCT GGA          480                                                                          Tyr Ala Phe Thr Asp Tyr Trp Leu Gly Trp Va - #l Lys His Arg Pro Gly           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - CAT GGA CTT GAG TGG ATT GGA GAT ATT TAC CC - #T GGA AGT GAT AAT ACT          528                                                                          His Gly Leu Glu Trp Ile Gly Asp Ile Tyr Pr - #o Gly Ser Asp Asn Thr           #               175                                                           - TAC TAC AAT GAG AAA TTC AAG GGC AAA GCC AC - #A CTG ACT ACA GAC AAA          576                                                                          Tyr Tyr Asn Glu Lys Phe Lys Gly Lys Ala Th - #r Leu Thr Thr Asp Lys           #           190                                                               - TCC TCG AGC ACA GCC TAT ATG CAG CTC AGT AG - #C CTG ACA TCT GAG GAC          624                                                                          Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Se - #r Leu Thr Ser Glu Asp           #       205                                                                   - TCT GCT GTC TAT TTC TGT GCA AGG GGC CTT AA - #A GGA GAC TAC TGG GGC          672                                                                          Ser Ala Val Tyr Phe Cys Ala Arg Gly Leu Ly - #s Gly Asp Tyr Trp Gly           #   220                                                                       - CAA GGG ACC ACC GTC ACC GTC TCC TCA GGT GG - #A GGC GGT TCA GGC GGA          720                                                                          Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gl - #y Gly Gly Ser Gly Gly           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GGT GGC TCT GGC GGT GGC GGA TCG GAC ATC CA - #G ATG ACC CAG TCT CCA          768                                                                          Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gl - #n Met Thr Gln Ser Pro           #               255                                                           - TCC TCA CTG TCT GCA TCT CTG GGA GGC AAA GT - #C ACC ATC ACT TGC AAG          816                                                                          Ser Ser Leu Ser Ala Ser Leu Gly Gly Lys Va - #l Thr Ile Thr Cys Lys           #           270                                                               - GCA AGC CAA GAC ATT AAG AAG TCT ATA GCT TG - #G TAC CAA CAC AAG CCT          864                                                                          Ala Ser Gln Asp Ile Lys Lys Ser Ile Ala Tr - #p Tyr Gln His Lys Pro           #       285                                                                   - GGA AAA GGT CCT AGG CTG CTC ATT CAT TAC AC - #A TCT ACA TTA CAG CCA          912                                                                          Gly Lys Gly Pro Arg Leu Leu Ile His Tyr Th - #r Ser Thr Leu Gln Pro           #   300                                                                       - GGC ATC CCA TCA AGG TTC AGT GGA AGT GGG TC - #T GGT GAA GAA TAT TCC          960                                                                          Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Se - #r Gly Glu Glu Tyr Ser           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - TTC AGC ATC AGC AAC CTG GAG CCT GAA GAT AT - #T GCA ACT TAT TAT TGT         1008                                                                          Phe Ser Ile Ser Asn Leu Glu Pro Glu Asp Il - #e Ala Thr Tyr Tyr Cys           #               335                                                           - CAA CAG TAT GAT AAT CTT CGG ACG TTC GGT GG - #A GGC ACC AAG CTG GAG         1056                                                                          Gln Gln Tyr Asp Asn Leu Arg Thr Phe Gly Gl - #y Gly Thr Lys Leu Glu           #           350                                                               #       1065                                                                  Leu Lys Arg                                                                           355                                                                   - (2) INFORMATION FOR SEQ ID NO:57:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 355 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala         #           110                                                               - Gln Ala Pro Lys Ser Asp Gln Val Gln Leu Gl - #n Gln Ser Gly Thr Glu         #       125                                                                   - Leu Ile Arg Pro Gly Thr Ser Val Lys Ile Se - #r Cys Lys Ala Ser Gly         #   140                                                                       - Tyr Ala Phe Thr Asp Tyr Trp Leu Gly Trp Va - #l Lys His Arg Pro Gly         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - His Gly Leu Glu Trp Ile Gly Asp Ile Tyr Pr - #o Gly Ser Asp Asn Thr         #               175                                                           - Tyr Tyr Asn Glu Lys Phe Lys Gly Lys Ala Th - #r Leu Thr Thr Asp Lys         #           190                                                               - Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Se - #r Leu Thr Ser Glu Asp         #       205                                                                   - Ser Ala Val Tyr Phe Cys Ala Arg Gly Leu Ly - #s Gly Asp Tyr Trp Gly         #   220                                                                       - Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gl - #y Gly Gly Ser Gly Gly         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gl - #n Met Thr Gln Ser Pro         #               255                                                           - Ser Ser Leu Ser Ala Ser Leu Gly Gly Lys Va - #l Thr Ile Thr Cys Lys         #           270                                                               - Ala Ser Gln Asp Ile Lys Lys Ser Ile Ala Tr - #p Tyr Gln His Lys Pro         #       285                                                                   - Gly Lys Gly Pro Arg Leu Leu Ile His Tyr Th - #r Ser Thr Leu Gln Pro         #   300                                                                       - Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Se - #r Gly Glu Glu Tyr Ser         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Phe Ser Ile Ser Asn Leu Glu Pro Glu Asp Il - #e Ala Thr Tyr Tyr Cys         #               335                                                           - Gln Gln Tyr Asp Asn Leu Arg Thr Phe Gly Gl - #y Gly Thr Lys Leu Glu         #           350                                                               - Leu Lys Arg                                                                         355                                                                   - (2) INFORMATION FOR SEQ ID NO:58:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 753 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..753                                                #/note= "IL2FBMetSerOnc"RMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                - GCA CCT ACT TCA ACT TCT ACA AAG AAA ACA CA - #G CTA CAA CTG GAG CAT           48                                                                          Ala Pro Thr Ser Thr Ser Thr Lys Lys Thr Gl - #n Leu Gln Leu Glu His           #                 15                                                          - TTA CTG CTG GAT TTA CAG ATG ATT TTG AAT GG - #A ATT AAT AAT TAC AAG           96                                                                          Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gl - #y Ile Asn Asn Tyr Lys           #             30                                                              - AAT CCC AAA CTC ACC AGG ATG CTC ACA TTT AA - #G TTT TAC ATG CCC AAG          144                                                                          Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Ly - #s Phe Tyr Met Pro Lys           #         45                                                                  - AAG GCC ACA GAA CTG AAA CAT CTT CAG TGT CT - #A GAA GAA GAA CTC AAA          192                                                                          Lys Ala Thr Glu Leu Lys His Leu Gln Cys Le - #u Glu Glu Glu Leu Lys           #     60                                                                      - CCT CTG GAG GAA GTG CTA AAT TTA GCT CAA AG - #C AAA AAC TTT CAC TTA          240                                                                          Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Se - #r Lys Asn Phe His Leu           # 80                                                                          - AGA CCC AGG GAC TTA ATC AGC AAT ATC AAC GT - #A ATA GTT CTG GAA CTA          288                                                                          Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Va - #l Ile Val Leu Glu Leu           #                 95                                                          - AAG GGA TCT GAA ACA ACA TTC ATG TGT GAA TA - #T GCT GAT GAG ACA GCA          336                                                                          Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Ty - #r Ala Asp Glu Thr Ala           #           110                                                               - ACC ATT GTA GAA TTT CTG AAC AGA TGG ATT AC - #C TTT TGT CAA AGC ATC          384                                                                          Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Th - #r Phe Cys Gln Ser Ile           #       125                                                                   - ATC TCA ACA CTG ACT GCC AAG AAA CTG AAC GA - #C GCT CAG GCG CCG AAG          432                                                                          Ile Ser Thr Leu Thr Ala Lys Lys Leu Asn As - #p Ala Gln Ala Pro Lys           #   140                                                                       - AGT GAT ATG TCA GAT TGG CTT ACA TTT CAG AA - #A AAA CAC ATC ACA AAC          480                                                                          Ser Asp Met Ser Asp Trp Leu Thr Phe Gln Ly - #s Lys His Ile Thr Asn           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - ACA AGG GAT GTT GAT TGT GAT AAT ATC ATG TC - #A ACA AAC TTG TTC CAC          528                                                                          Thr Arg Asp Val Asp Cys Asp Asn Ile Met Se - #r Thr Asn Leu Phe His           #               175                                                           - TGC AAG GAC AAG AAC ACT TTT ATC TAT TCA CG - #T CCT GAG CCA GTG AAG          576                                                                          Cys Lys Asp Lys Asn Thr Phe Ile Tyr Ser Ar - #g Pro Glu Pro Val Lys           #           190                                                               - GCC ATC TGT AAA GGA ATT ATA GCC TCC AAA AA - #T GTG TTA ACT ACC TCT          624                                                                          Ala Ile Cys Lys Gly Ile Ile Ala Ser Lys As - #n Val Leu Thr Thr Ser           #       205                                                                   - GAG TTT TAT CTC TCT GAT TGC AAT GTA ACA AG - #C AGG CCT TGC AAG TAT          672                                                                          Glu Phe Tyr Leu Ser Asp Cys Asn Val Thr Se - #r Arg Pro Cys Lys Tyr           #   220                                                                       - AAA TTA AAG AAA TCA ACT AAT AAA TTT TGT GT - #A ACT TGT GAA AAT CAG          720                                                                          Lys Leu Lys Lys Ser Thr Asn Lys Phe Cys Va - #l Thr Cys Glu Asn Gln           225                 2 - #30                 2 - #35                 2 -       #40                                                                           #        753T CAT TTT GTT GGA GTT GGA TCT TG - #T                             Ala Pro Val His Phe Val Gly Val Gly Ser Cy - #s                               #               250                                                           - (2) INFORMATION FOR SEQ ID NO:59:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 251 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                - Ala Pro Thr Ser Thr Ser Thr Lys Lys Thr Gl - #n Leu Gln Leu Glu His         #                 15                                                          - Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gl - #y Ile Asn Asn Tyr Lys         #             30                                                              - Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Ly - #s Phe Tyr Met Pro Lys         #         45                                                                  - Lys Ala Thr Glu Leu Lys His Leu Gln Cys Le - #u Glu Glu Glu Leu Lys         #     60                                                                      - Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Se - #r Lys Asn Phe His Leu         # 80                                                                          - Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Va - #l Ile Val Leu Glu Leu         #                 95                                                          - Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Ty - #r Ala Asp Glu Thr Ala         #           110                                                               - Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Th - #r Phe Cys Gln Ser Ile         #       125                                                                   - Ile Ser Thr Leu Thr Ala Lys Lys Leu Asn As - #p Ala Gln Ala Pro Lys         #   140                                                                       - Ser Asp Met Ser Asp Trp Leu Thr Phe Gln Ly - #s Lys His Ile Thr Asn         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Thr Arg Asp Val Asp Cys Asp Asn Ile Met Se - #r Thr Asn Leu Phe His         #               175                                                           - Cys Lys Asp Lys Asn Thr Phe Ile Tyr Ser Ar - #g Pro Glu Pro Val Lys         #           190                                                               - Ala Ile Cys Lys Gly Ile Ile Ala Ser Lys As - #n Val Leu Thr Thr Ser         #       205                                                                   - Glu Phe Tyr Leu Ser Asp Cys Asn Val Thr Se - #r Arg Pro Cys Lys Tyr         #   220                                                                       - Lys Leu Lys Lys Ser Thr Asn Lys Phe Cys Va - #l Thr Cys Glu Asn Gln         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Ala Pro Val His Phe Val Gly Val Gly Ser Cy - #s                             #               250                                                           - (2) INFORMATION FOR SEQ ID NO:60:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 768 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..765                                                #/note= "MetSerOncFBIL2"RMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                - ATG TCA GAT TGG CTT ACA TTT CAG AAA AAA CA - #C ATC ACA AAC ACA AGG           48                                                                          Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg           #                 15                                                          - GAT GTT GAT TGT GAT AAT ATC ATG TCA ACA AA - #C TTG TTC CAC TGC AAG           96                                                                          Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys           #             30                                                              - GAC AAG AAC ACT TTT ATC TAT TCA CGT CCT GA - #G CCA GTG AAG GCC ATC          144                                                                          Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile           #         45                                                                  - TGT AAA GGA ATT ATA GCC TCC AAA AAT GTG TT - #A ACT ACC TCT GAG TTT          192                                                                          Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe           #     60                                                                      - TAT CTC TCT GAT TGC AAT GTA ACA AGC AGG CC - #T TGC AAG TAT AAA TTA          240                                                                          Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu           # 80                                                                          - AAG AAA TCA ACT AAT AAA TTT TGT GTA ACT TG - #T GAA AAT CAG GCA CCA          288                                                                          Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro           #                 95                                                          - GTT CAT TTT GTT GGA GTT GGA TCT TGT GCC AA - #G AAA CTG AAC GAC GCT          336                                                                          Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala           #           110                                                               - CAG GCG CCG AAG AGT GAT GCA CCT ACT TCA AC - #T TCT ACA AAG AAA ACA          384                                                                          Gln Ala Pro Lys Ser Asp Ala Pro Thr Ser Th - #r Ser Thr Lys Lys Thr           #       125                                                                   - CAG CTA CAA CTG GAG CAT TTA CTG CTG GAT TT - #A CAG ATG ATT TTG AAT          432                                                                          Gln Leu Gln Leu Glu His Leu Leu Leu Asp Le - #u Gln Met Ile Leu Asn           #   140                                                                       - GGA ATT AAT AAT TAC AAG AAT CCC AAA CTC AC - #C AGG ATG CTC ACA TTT          480                                                                          Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Th - #r Arg Met Leu Thr Phe           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - AAG TTT TAC ATG CCC AAG AAG GCC ACA GAA CT - #G AAA CAT CTT CAG TGT          528                                                                          Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Le - #u Lys His Leu Gln Cys           #               175                                                           - CTA GAA GAA GAA CTC AAA CCT CTG GAG GAA GT - #G CTA AAT TTA GCT CAA          576                                                                          Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Va - #l Leu Asn Leu Ala Gln           #           190                                                               - AGC AAA AAC TTT CAC TTA AGA CCC AGG GAC TT - #A ATC AGC AAT ATC AAC          624                                                                          Ser Lys Asn Phe His Leu Arg Pro Arg Asp Le - #u Ile Ser Asn Ile Asn           #       205                                                                   - GTA ATA GTT CTG GAA CTA AAG GGA TCT GAA AC - #A ACA TTC ATG TGT GAA          672                                                                          Val Ile Val Leu Glu Leu Lys Gly Ser Glu Th - #r Thr Phe Met Cys Glu           #   220                                                                       - TAT GCT GAT GAG ACA GCA ACC ATT GTA GAA TT - #T CTG AAC AGA TGG ATT          720                                                                          Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Ph - #e Leu Asn Arg Trp Ile           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - ACC TTT TGT CAA AGC ATC ATC TCA ACA CTG AC - #T CAT CAC CAT                 # 762                                                                         Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Th - #r His His His                   #               250                                                           #          768                                                                - (2) INFORMATION FOR SEQ ID NO:61:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 254 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                - Met Ser Asp Trp Leu Thr Phe Gln Lys Lys Hi - #s Ile Thr Asn Thr Arg         #                 15                                                          - Asp Val Asp Cys Asp Asn Ile Met Ser Thr As - #n Leu Phe His Cys Lys         #             30                                                              - Asp Lys Asn Thr Phe Ile Tyr Ser Arg Pro Gl - #u Pro Val Lys Ala Ile         #         45                                                                  - Cys Lys Gly Ile Ile Ala Ser Lys Asn Val Le - #u Thr Thr Ser Glu Phe         #     60                                                                      - Tyr Leu Ser Asp Cys Asn Val Thr Ser Arg Pr - #o Cys Lys Tyr Lys Leu         # 80                                                                          - Lys Lys Ser Thr Asn Lys Phe Cys Val Thr Cy - #s Glu Asn Gln Ala Pro         #                 95                                                          - Val His Phe Val Gly Val Gly Ser Cys Ala Ly - #s Lys Leu Asn Asp Ala         #           110                                                               - Gln Ala Pro Lys Ser Asp Ala Pro Thr Ser Th - #r Ser Thr Lys Lys Thr         #       125                                                                   - Gln Leu Gln Leu Glu His Leu Leu Leu Asp Le - #u Gln Met Ile Leu Asn         #   140                                                                       - Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Th - #r Arg Met Leu Thr Phe         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Le - #u Lys His Leu Gln Cys         #               175                                                           - Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Va - #l Leu Asn Leu Ala Gln         #           190                                                               - Ser Lys Asn Phe His Leu Arg Pro Arg Asp Le - #u Ile Ser Asn Ile Asn         #       205                                                                   - Val Ile Val Leu Glu Leu Lys Gly Ser Glu Th - #r Thr Phe Met Cys Glu         #   220                                                                       - Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Ph - #e Leu Asn Arg Trp Ile         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Th - #r His His His                 #               250                                                           - (2) INFORMATION FOR SEQ ID NO:62:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 387 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: DNA                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..387                                                #/note= "SigPepOnc" INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                - ATG GGT CTG GAA AAA TCT CTG ATC CTG TTC CC - #G CTG TTC TTC CTG CTG           48                                                                          Met Gly Leu Glu Lys Ser Leu Ile Leu Phe Pr - #o Leu Phe Phe Leu Leu           #                 15                                                          - CTG GGT TGG GTT CAG CCG TCT CTG GGT CAG GA - #T TGG CTT ACA TTT CAG           96                                                                          Leu Gly Trp Val Gln Pro Ser Leu Gly Gln As - #p Trp Leu Thr Phe Gln           #             30                                                              - AAA AAA CAC ATC ACA AAC ACA AGG GAT GTT GA - #T TGT GAT AAT ATC ATG          144                                                                          Lys Lys His Ile Thr Asn Thr Arg Asp Val As - #p Cys Asp Asn Ile Met           #         45                                                                  - TCA ACA AAC TTG TTC CAC TGC AAG GAC AAG AA - #C ACT TTT ATC TAT TCA          192                                                                          Ser Thr Asn Leu Phe His Cys Lys Asp Lys As - #n Thr Phe Ile Tyr Ser           #     60                                                                      - CGT CCT GAG CCA GTG AAG GCC ATC TGT AAA GG - #A ATT ATA GCC TCC AAA          240                                                                          Arg Pro Glu Pro Val Lys Ala Ile Cys Lys Gl - #y Ile Ile Ala Ser Lys           # 80                                                                          - AAT GTG TTA ACT ACC TCT GAG TTT TAT CTC TC - #T GAT TGC AAT GTA ACA          288                                                                          Asn Val Leu Thr Thr Ser Glu Phe Tyr Leu Se - #r Asp Cys Asn Val Thr           #                 95                                                          - AGC AGG CCT TGC AAG TAT AAA TTA AAG AAA TC - #A ACT AAT AAA TTT TGT          336                                                                          Ser Arg Pro Cys Lys Tyr Lys Leu Lys Lys Se - #r Thr Asn Lys Phe Cys           #           110                                                               - GTA ACT TGT GAA AAT CAG GCA CCA GTT CAT TT - #T GTT GGA GTT GGA TCT          384                                                                          Val Thr Cys Glu Asn Gln Ala Pro Val His Ph - #e Val Gly Val Gly Ser           #       125                                                                   #            387                                                              Cys                                                                           - (2) INFORMATION FOR SEQ ID NO:63:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 129 amino                                                         (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                - Met Gly Leu Glu Lys Ser Leu Ile Leu Phe Pr - #o Leu Phe Phe Leu Leu         #                 15                                                          - Leu Gly Trp Val Gln Pro Ser Leu Gly Gln As - #p Trp Leu Thr Phe Gln         #             30                                                              - Lys Lys His Ile Thr Asn Thr Arg Asp Val As - #p Cys Asp Asn Ile Met         #         45                                                                  - Ser Thr Asn Leu Phe His Cys Lys Asp Lys As - #n Thr Phe Ile Tyr Ser         #     60                                                                      - Arg Pro Glu Pro Val Lys Ala Ile Cys Lys Gl - #y Ile Ile Ala Ser Lys         # 80                                                                          - Asn Val Leu Thr Thr Ser Glu Phe Tyr Leu Se - #r Asp Cys Asn Val Thr         #                 95                                                          - Ser Arg Pro Cys Lys Tyr Lys Leu Lys Lys Se - #r Thr Asn Lys Phe Cys         #           110                                                               - Val Thr Cys Glu Asn Gln Ala Pro Val His Ph - #e Val Gly Val Gly Ser         #       125                                                                   - Cys                                                                         - (2) INFORMATION FOR SEQ ID NO:64:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 355 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (ix) FEATURE:                                                                     (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..355                                                #/note= "E6FB[Met-(-1)]SerrOnc":                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                - Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Me - #t Tyr Ala Ser Leu Gly         #                15                                                           - Glu Arg Val Thr Phe Thr Cys Lys Ala Ser Gl - #n Asp Ile Asn Asn Tyr         #            30                                                               - Leu Cys Trp Phe Gln Gln Lys Pro Gly Lys Se - #r Pro Lys Thr Leu Ile         #        45                                                                   - Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pr - #o Ser Arg Phe Ser Gly         #    60                                                                       - Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Il - #e Ser Ser Leu Glu Tyr         #80                                                                           - Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Ty - #r Asp Glu Phe Pro Tyr         #                95                                                           - Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Ly - #s Gly Gly Gly Gly Ser         #           110                                                               - Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #u Val Gln Leu Gln Gln         #       125                                                                   - Ser Gly Thr Val Leu Ala Arg Pro Gly Ala Se - #r Val Lys Met Ser Cys         #   140                                                                       - Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Tr - #p Met His Trp Ile Lys         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile Va - #l Ala Ile Asp Pro Arg         #               175                                                           - Asn Ser Asp Thr Ile Tyr Asn Pro Gln Phe Ly - #s His Lys Ala Lys Leu         #           190                                                               - Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr Me - #t Glu Leu Asn Ser Leu         #       205                                                                   - Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys Th - #r Pro Leu Tyr Tyr Phe         #   220                                                                       - Asp Ser Trp Gly Gln Gly Thr Thr Leu Thr Va - #l Ser Ser Ala Lys Lys         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Leu Asn Asp Ala Gln Ala Pro Lys Ser Asp Me - #t Ser Asp Trp Leu Thr         #               255                                                           - Phe Gln Lys Lys His Ile Thr Asn Thr Arg As - #p Val Asp Cys Asp Asn         #           270                                                               - Ile Met Ser Thr Asn Leu Phe His Cys Lys As - #p Lys Asn Thr Phe Ile         #       285                                                                   - Tyr Ser Arg Pro Glu Pro Val Lys Ala Ile Cy - #s Lys Gly Ile Ile Ala         #   300                                                                       - Ser Lys Asn Val Leu Thr Thr Ser Glu Phe Ty - #r Leu Ser Asp Cys Asn         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Val Thr Ser Arg Pro Cys Lys Tyr Lys Leu Ly - #s Lys Ser Thr Asn Lys         #               335                                                           - Phe Cys Val Thr Cys Glu Asn Gln Ala Pro Va - #l His Phe Val Gly Val         #           350                                                               - Gly Ser Cys                                                                         355                                                                   __________________________________________________________________________

What is claimed is:
 1. A ribonuclease molecule comprising: (a) an aminoterminal end beginning with a methionine which is followed by any aminoacid other than glutamic acid; (b) when aligned for maximumcorrespondence with SEQ ID NO:13, a cysteine at amino acid positions 26,40, 58, 84, 95 and 110; a lysine at position 41 and a histidine atposition 119, and (c) an nOnc-derived amino acid sequence; wherein saidribonuclease molecule has measurable ribonuclease activity.
 2. Theribonuclease of claim 1 which has an amino terminal end selected fromthe group consisting of: Met-Lys; Met-Tyr; Met-Ser; Met-Ala; Met-Arg;and Met-Asn.
 3. The ribonuclease of claim 1, which has an amino terminalend selected from the group consisting of:Met-Ala; Met-Ala-Ala;Met-Ala-Ala-Ser; Met-Arg; Met-(J); Met-Lys-(J); Met-Arg-(J); Met-Lys;Met-Lys-Pro; Met-Lys-(J)-Pro (SEQ ID NO:14); Met-Lys-Pro-(J) (SEQ IDNO:15); Met-Asn; Met-Gln; Met-Asn-(J); Met-Gln-(J); Met-Asn-(J)-Pro (SEQID NO:16); Met-(J)-Lys; Met-(J)-Lys-Pro (SEQ ID NO:17); andMet-(J)-Pro-Lys (SEQ ID NO:18);where (J) is Ser, Tyr or Thr.
 4. Theribonuclease of claim 1, which has an amino terminal end of Met-Ala. 5.The ribonuclease of claim 1, which has an amino terminal end of Met-Arg.6. The ribonuclease of claim 1, which has an amino terminal end ofMet-Lys.
 7. The ribonuclease of claim 1, which has an amino terminal endof Met-Asn.
 8. The ribonuclease of claim 1, which has an amino terminalend of Met-Gln.
 9. The ribonuclease of claim 1, which has an aminoterminal end selected from the group consisting of Met-Ser; Met-Tyr orMet-Thr.
 10. The ribonuclease of claim 3, wherein aspartic acid of aminoacid position 2 of the amino acid sequence of (c) or position 4 withreference to the sequence of bovine RNase is deleted or replaced by Alaor Asn.
 11. The ribonuclease of claim 1 wherein the amino acid sequencecomprises a sequence having the formula:

    Met(-1) eosinophil derived neurotoxin.sub.(1-m) Onc.sub.(n-104)

wherein Met(-1) refers to an amino terminal residue of Met; whereineosinophil derived neurotoxin.sub.(1-m) refers to a contiguous sequenceof amino acids of a length beginning at amino acid position 1 ofeosinophil derived neurotoxin (SEQ ID NO:9) and continuing to andincluding amino acid position "m" of eosinophil derived neurotoxin;wherein Onc.sub.(n-104) refers to a sequence of contiguous amino acidsbeginning at no acid position "n" and continuing to and including aminoacid position 104 as set out in SEQ ID NO:1; and wherein "m" is theamino acid position of eosinophil derived neurotoxin selected from thegroup consisting of 5, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22; suchthat: when m is 21, n is 16 or 17; when m is 22, n is 17; when m is 20,n is 16; when m is 19, n is 15; when m is 18, n is 14; when m is 17, nis 12 or 13; when m is 16, n is 11, 12, 13 or 14; when m is 15, n is 10;when m is 14,n is9; when m is 13, n is 8; and when m is 5, n is
 1. 12.The ribonuclease of claim 1, comprising an amino acid sequencesubstantially identical to that of SEQ ID NO:28.
 13. The ribonuclease ofclaim 1, comprising an amino acid sequence substantially identical tothat of SEQ ID NO:22.
 14. The ribonuclease of claim 1, comprising anamino acid sequence substantially identical to that of SEQ ID NO:24. 15.The ribonuclease of claim 1, comprising an amino acid sequencesubstantially identical to that of SEQ ID NO:26.
 16. The ribonuclease ofclaim 1, comprising an amino acid sequence substantially identical tothat of SEQ ID NO:30.
 17. The ribonuclease of claim 1, comprising anamino acid sequence substantially identical to that of SEQ ID NO:32. 18.The ribonuclease of claim 1, which includes an amino acid sequencesubstantially identical to that of SEQ ID NO:2.
 19. The ribonuclease ofclaim 1, comprising a carboxyl terminal end derived from angiogenincorresponding to the amino acid sequence of positions 101 to 107 of SEQID NO:20.
 20. The ribonuclease of claim 19, comprising an amino acidsequence substantially identical to that of SEQ ID NO:20.
 21. Anisolated amino acid sequence substantially identical to that set out inSEQ ID NO:2.
 22. A fusion protein comprising the ribonuclease of claim 1joined to a ligand binding moiety or label.
 23. The fusion protein ofclaim 22, further comprising an antibody.
 24. An isolated nucleic acidsequence encoding the amino acid sequence of claim
 1. 25. Apharmaceutical composition comprising a cytotoxic amount of aribonuclease of claim 1 and a pharmaceutically acceptable carrier. 26.The pharmaceutical composition of claim 25 wherein the ribonuclease isjoined to a ligand binding moiety.
 27. A method of selectively killingcells comprising contacting cells to be killed with a ribonuclease ofclaim 1 joined to a ligand binding moiety.
 28. The ribonuclease moleculeof claim 1 which further has a nuclear localization signal.
 29. Theribonuclease molecule of claim 1 which further has an endoplasmicretention sequence.
 30. A vector comprising a nucleic acid encoding aribonuclease of claim
 1. 31. A host cell comprising a nucleic acidencoding a ribonuclease of claim 1.